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Makoto Shimoyama - One of the best experts on this subject based on the ideXlab platform.
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Exocytosis of arginine-specific ADP-ribosyltransferase and P33 induced by A23187 and calcium or serum-opsonized zymosan in chicken polymorphonuclear leukocytes.
Journal of biochemistry, 1996Co-Authors: Masaharu Terashima, Mikako Tsuchiya, Muhammad Badruzzaman, Makoto ShimoyamaAbstract:Exocytosis is a common phenomenon in neutrophil functions. We earlier reported the co-localization of arginine-specific ADP-ribosyltransferase [EC 2.4.2.31] and its target Protein P33 (mim-1 Protein) in cytoplasmic granules in chicken polymorphonuclear leukocytes (so-called heterophils) [Mishima, K., Terashima, M., Obara, S., Yamada, K., Imai, K., and Shimoyama, M. (1991) J. Biochem. 110, 388-394]. In the present study, we obtained evidence that the transferase and P33 were released into the extracellular space by the stimulus of calcium ionophore A23187 or serum-opsonized zymosan, but scarcely by phorbol myristate acetate (PMA) or N-formyl-Met-Leu-Phe (fMLP), thereby indicating the co-localization of the transferase and P33 in the azurophilic granules, and not in specific granules. [32P]ADP-ribosylation of P33 occurred in the extracellular space, induced by the stimulus of A23187 or opsonized zymosan in the presence of [32P]NAD. Our findings are interpreted to mean that heterophil transferase and P33 may be involved in neutrophil functions during processes of inflammation.
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Target Protein for eucaryotic arginine-specific ADP-ribosyltransferase
Molecular and Cellular Biochemistry, 1994Co-Authors: Mikako Tsuchiya, Makoto ShimoyamaAbstract:Among ADP-ribosyltransferases reported in eucaryotes, arginine-specific transferases from turkey erythrocytes, chicken heterophils and rabbit skeletal muscle have been purified and extensively studied. They were reported to modify a number of Proteins in vitro . ADP-ribosylation of Ha- ras -p21 and transducin by the turkey erythrocyte transferase inhibits their GTPase and GTP-binding activities. Chicken heterophil enzyme modifies several substrate Proteins for Protein kinases and decreases the phosphate-acceptor activity. Rabbit skeletal muscle Ca^2+-ATPase is inhibited by ADP-ribosylation catalyzed by the muscle transferase. Three transferases all ADP-ribosylate small molecular weight guanidino compounds such as arginine, arginine methylester and agmatine and poly-L-arginine and nuclear histones. However, the observation that muscle transferase did not ADP-ribosylate casein or actin, both of which can be modified by the heterophil transferase under the same conditions indicates that substrate specificity of these two enzymes are different. Substrate-dependent effects were observed with polyions of nucleotides such that polyanions stimulate the ADP-ribosylation of possible target Protein, P33 by chicken heterophil transferase but has no effect on the modification of casein by the same enzyme.
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Target Protein for eucaryotic arginine-specific ADP-ribosyltransferase
Molecular and Cellular Biochemistry, 1994Co-Authors: Mikako Tsuchiya, Makoto ShimoyamaAbstract:Among ADP-ribosyltransferases reported in eucaryotes, arginine-specific transferases from turkey erythrocytes, chicken heterophils and rabbit skeletal muscle have been purified and extensively studied. They were reported to modify a number of Proteins in vitro. ADP-ribosylation of Ha-ras-p21 and transducin by the turkey erythrocyte transferase inhibits their GTPase and GTP-binding activities. Chicken heterophil enzyme modifies several substrate Proteins for Protein kinases and decreases the phosphate-acceptor activity. Rabbit skeletal muscle Ca2+-ATPase is inhibited by ADP-ribosylation catalyzed by the muscle transferase. Three transferases all ADP-ribosylate small molecular weight guanidino compounds such as arginine, arginine methylester and agmatine and poly-L-arginine and nuclear histones. However, the observation that muscle transferase did not ADP-ribosylate casein or actin, both of which can be modified by the heterophil transferase under the same conditions indicates that substrate specificity of these two enzymes are different. Substrate-dependent effects were observed with polyions of nucleotides such that polyanions stimulate the ADP-ribosylation of possible target Protein, P33 by chicken heterophil transferase but has no effect on the modification of casein by the same enzyme. (Mol Cell Biochem 138: 113–118, 1994)
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adp ribosylation of actins by arginine specific adp ribosyltransferase purified from chicken heterophils
FEBS Journal, 1992Co-Authors: Masaharu Terashima, Koichi Mishima, Kazuo Yamada, Mikako Tsuchiya, Tadao Wakutani, Makoto ShimoyamaAbstract:We reported the purification and characterization of an arginine-specific ADP-ribosyltransferase and acceptor Protein P33 in granules of chicken peripheral polymorphonuclear leukocytes (heterophils) [Mishima, K., Terashima, M., Obara, S., Yamada, K., Imai, K. & Shimoyama, M. (1991) J. Biochem. (Tokyo) 110, 388–394]. In the present study, we obtained evidence that chicken non-muscle β/γ-actin, skeletal muscle α-actin and smooth-muscle γ-actin were ADP ribosylated by the heterophil ADP-ribosyltransferase. The stoichiometry of ADP–ribose incorporation into these actins was 1.2 mol, 1.0 mol and 2.0 mol ADP–ribose/mol of β/γ-actin, α-actin and γ-actin, respectively. The optimal pH for the ADP ribosylation was at pH 8.5, with the respective actin. Km values for NAD were calculated to be 30 μM with β/γ-actin, 35 μM with α-actin and 20 μM with γ-actin. The Km values for the actin isoforms were 15 μM for β/γ-actin, 2.5 μM for α-actin and 10 μM for γ-actin. ADP ribosylation of actin inhibited its capacity to polymerize, as determined by the increase in fluoresence intensity with N-(1-pyrenyl)iodoacetamide-labelled actin. Filamentous actin (F-actin) polymerized with the respective actin isoform was also ADP ribosylated, although the extent of the modification of F-actin was lower than that of globular actin (G-actin). In situ ADP ribosylation of β/γ-actin was evidenced with chicken peripheral heterophils permeabilized with saponin. Thus, the endogenous ADP ribosylation of actin in the heterophils may be involved in the cellular processes such as phagocytosis, secretion and migration.
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Arginine-specific ADP-ribosyltransferase and its acceptor Protein P33 in chicken polymorphonuclear cells: co-localization in the cell granules, partial characterization, and in situ mono(ADP-ribosyl)ation.
Journal of biochemistry, 1991Co-Authors: Koichi Mishima, Masaharu Terashima, Kazuo Yamada, Seiji Obara, Katsuyuki Imai, Makoto ShimoyamaAbstract:We have reported the purification and characterization of arginine-specific ADP-ribosyltransferase from hen liver nuclei [Tanigawa, Y. et al. (1984) J. Biol. Chem. 259, 2022-2029] and the DNA-dependent mono(ADP-ribosyl)ation of P33, an acceptor Protein in the nuclei [Mishima, K. et al. (1989) Eur. J. Biochem. 179, 267-273]. In the present study, we obtained evidence that among various tissues and cells from chicken, polymorphonuclear cells, so-called heterophils, possess both the ADP-ribosyltransferase and P33 at high levels. Percoll density gradient centrifugation of the postnuclear fraction of the heterophils revealed the co-localization of ADP-ribosyltransferase with P33 in the granule fraction. The enzyme and P33 were purified approximately 219- and 3.77-fold, respectively, from postnuclear pellet fraction to apparent homogeneity. The properties of heterophil ADP-ribosyltransferase and P33 were compared with those of the liver enzyme and P33. The molecular mass of the heterophil enzyme was estimated by SDS-polyacrylamide gel electrophoresis to be 27.5 kDa. The enzyme activity was stimulated by a sulfhydryl agent and inhibited by lysolecithin, NaCl, and inorganic phosphate. The mono(ADP-ribosyl)ation of P33 was markedly enhanced by polyanion, such as DNA, RNA, or poly(L-glutamate). SDS-polyacrylamide gel electrophoretic analysis after limited trypsin proteolysis of P33s, purified from chicken heterophils and liver, showed much the same pattern. Thus, it appears that ADP-ribosyltransferase and P33 present in heterophils are identical to those in the liver, respectively. P33 is considered to be an in situ substrate for ADP-ribosyltransferase, since it was specifically mono(ADP-ribosyl)ated in permeabilized heterophils.(ABSTRACT TRUNCATED AT 250 WORDS)
Kyoko Kojima - One of the best experts on this subject based on the ideXlab platform.
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Characterization of Carbohydrate-binding Protein P33/41: RELATION WITH ANNEXIN IV, MOLECULAR BASIS OF THE DOUBLET FORMS (P33 AND p41), AND MODULATION OF THE CARBOHYDRATE BINDING ACTIVITY BY PHOSPHOLIPIDS (∗)
The Journal of biological chemistry, 1996Co-Authors: Kyoko Kojima, Kazuo Yamamoto, Tatsuro Irimura, Toshiaki Osawa, Haruko Ogawa, Isamu MatsumotoAbstract:A Protein, P33/41, expressed in bovine kidney and many other tissues was identified as a lectin which binds to sialoglycoProteins and glycosaminoglycans in a calcium-dependent manner. Partial amino acid sequences of P33/41 are highly homologous to those of a calcium/phospholipid-binding annexin Protein, annexin IV (endonexin). P33/41 exhibited similar calcium-dependent phospholipid binding activity (Kojima, K., Ogawa, H., Seno, N., Yamamoto, K., Irimura, T., Osawa, T., and Matsumoto, I. (1992) J. Biol. Chem. 267, 20536-20539). To further characterize P33/41, we cloned the P33/41 cDNA and characterized the recombinant Protein encoded by this cDNA. Oligonucleotide probes were synthesized based on partial amino acid sequences of P33/41 and used for screening. A P33/41 cDNA clone was isolated encoding a Protein of 319 amino acids with a calculated molecular mass of 35,769 Da. The deduced amino acid sequence was identical to that of bovine annexin IV except for one amino acid substitution. The recombinant Protein gave two 33-kDa (P33) and 41-kDa (p41) bands on SDS-polyacrylamide gel electrophoresis under non-reducing conditions, and only one 33-kDa band under reducing conditions, as did the native Protein. Mass spectrometric analysis combined with site-directed mutagenesis of each of the four cysteine residues of the recombinant Protein revealed that p41 is a dimer of P33 cross-linked at Cys-198 via a disulfide bond. The recombinant Protein bound to columns of heparin and fetuin glycopeptides in a calcium dependent manner and to phospholipid vesicles composed of phosphatidylserine (PS)/phosphatidylcholine (PC), phosphatidylethanolamine (PE)/PC or phosphatidylinositol (PI)/PC. Furthermore, concurrent binding assays showed that the binding of the recombinant Protein to phospholipid vesicles was not affected by heparin, whereas that to heparin was influenced by the phospholipid composition of the vesicles; the highest binding was observed with vesicles composed of PE/PC. These results suggest that P33/41 binds two types of ligands via different sites and that phospholipids modulate the carbohydrate binding activity of P33/41.
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characterization of carbohydrate binding Protein P33 41 relation with annexin iv molecular basis of the doublet forms P33 and p41 and modulation of the carbohydrate binding activity by phospholipids
Journal of Biological Chemistry, 1996Co-Authors: Kyoko Kojima, Kazuo Yamamoto, Tatsuro Irimura, Toshiaki Osawa, Haruko Ogawa, Isamu MatsumotoAbstract:A Protein, P33/41, expressed in bovine kidney and many other tissues was identified as a lectin which binds to sialoglycoProteins and glycosaminoglycans in a calcium-dependent manner. Partial amino acid sequences of P33/41 are highly homologous to those of a calcium/phospholipid-binding annexin Protein, annexin IV (endonexin). P33/41 exhibited similar calcium-dependent phospholipid binding activity (Kojima, K., Ogawa, H., Seno, N., Yamamoto, K., Irimura, T., Osawa, T., and Matsumoto, I. (1992) J. Biol. Chem. 267, 20536-20539). To further characterize P33/41, we cloned the P33/41 cDNA and characterized the recombinant Protein encoded by this cDNA. Oligonucleotide probes were synthesized based on partial amino acid sequences of P33/41 and used for screening. A P33/41 cDNA clone was isolated encoding a Protein of 319 amino acids with a calculated molecular mass of 35,769 Da. The deduced amino acid sequence was identical to that of bovine annexin IV except for one amino acid substitution. The recombinant Protein gave two 33-kDa (P33) and 41-kDa (p41) bands on SDS-polyacrylamide gel electrophoresis under non-reducing conditions, and only one 33-kDa band under reducing conditions, as did the native Protein. Mass spectrometric analysis combined with site-directed mutagenesis of each of the four cysteine residues of the recombinant Protein revealed that p41 is a dimer of P33 cross-linked at Cys-198 via a disulfide bond. The recombinant Protein bound to columns of heparin and fetuin glycopeptides in a calcium dependent manner and to phospholipid vesicles composed of phosphatidylserine (PS)/phosphatidylcholine (PC), phosphatidylethanolamine (PE)/PC or phosphatidylinositol (PI)/PC. Furthermore, concurrent binding assays showed that the binding of the recombinant Protein to phospholipid vesicles was not affected by heparin, whereas that to heparin was influenced by the phospholipid composition of the vesicles; the highest binding was observed with vesicles composed of PE/PC. These results suggest that P33/41 binds two types of ligands via different sites and that phospholipids modulate the carbohydrate binding activity of P33/41.
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Expression of Carbohydrate-Binding Protein P33/41 in Human Tumor Cell Lines
Journal of biochemistry, 1996Co-Authors: Ayano Satoh, Kyoko Kojima, Haruko Ogawa, Eiji Takayama, Takao Yamori, Shigeo Sato, Tokuichi Kawaguchi, Takashi Tsuruo, Yoshimoto Katsura, Tatsuo KinaAbstract:We previously reported a new type of lectin, P33/41 (annexin IV), which was isolated from a bovine tissue extract [Kojima, K. et al. (1992) J. Biol. Chem. 267, 20536-20539]. When the expression of P33/41 (annexin IV) was surveyed in the lysates of 39 human tumor cell lines by SDS-PAGE, followed by Western blot analysis with polyclonal anti-bovine P33/41 and monoclonal anti-annexin IV (Z016, Zymed) antibodies, 21 cell lines were found to be reactive with the polyclonal antibody, whereas all 39 cell lines were stained with Z016. These results together with those obtained with standard Proteins, annexins IV and V, suggested that the monoclonal antibody, Z016, recognizes annexin V, but not P33/41 (annexin IV). Therefore, we performed cDNA cloning of human P33/41 (annexin IV) to prepare a recombinant Protein and raised monoclonal antibodies against the Protein. Northern blot analysis with the cDNA as a probe showed that a human colon cancer cell line, HT29, contains P33/41 (annexin IV) mRNA of two sizes, 2.0 and 3.0 kb. The two monoclonal antibodies, AS11 and AS17, against the recombinant Protein generated were useful for flow cytometric analysis, ELISA, Western blot analysis and immunoprecipitation. Flow cytometric analysis with AS17 showed that P33/41 (annexin IV) is located in the cytoplasm of HT29 cells, but not on the cell surface. However, one of the cell surface Proteins first labeled with biotin and then solubilized with a detergent was immunoprecipitated with AS17. The results suggest the existence of a membrane spanning form of P33/41 (annexin IV).
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expression of carbohydrate binding Protein P33 41 in human tumor cell lines
Journal of Biochemistry, 1996Co-Authors: Ayano Satoh, Kyoko Kojima, Haruko Ogawa, Eiji Takayama, Takao Yamori, Shigeo Sato, Tokuichi Kawaguchi, Takashi Tsuruo, Yoshimoto Katsura, Tatsuo KinaAbstract:We previously reported a new type of lectin, P33/41 (annexin IV), which was isolated from a bovine tissue extract [Kojima, K. et al. (1992) J. Biol. Chem. 267, 20536-20539]. When the expression of P33/41 (annexin IV) was surveyed in the lysates of 39 human tumor cell lines by SDS-PAGE, followed by Western blot analysis with polyclonal anti-bovine P33/41 and monoclonal anti-annexin IV (Z016, Zymed) antibodies, 21 cell lines were found to be reactive with the polyclonal antibody, whereas all 39 cell lines were stained with Z016. These results together with those obtained with standard Proteins, annexins IV and V, suggested that the monoclonal antibody, Z016, recognizes annexin V, but not P33/41 (annexin IV). Therefore, we performed cDNA cloning of human P33/41 (annexin IV) to prepare a recombinant Protein and raised monoclonal antibodies against the Protein. Northern blot analysis with the cDNA as a probe showed that a human colon cancer cell line, HT29, contains P33/41 (annexin IV) mRNA of two sizes, 2.0 and 3.0 kb. The two monoclonal antibodies, AS11 and AS17, against the recombinant Protein generated were useful for flow cytometric analysis, ELISA, Western blot analysis and immunoprecipitation. Flow cytometric analysis with AS17 showed that P33/41 (annexin IV) is located in the cytoplasm of HT29 cells, but not on the cell surface. However, one of the cell surface Proteins first labeled with biotin and then solubilized with a detergent was immunoprecipitated with AS17. The results suggest the existence of a membrane spanning form of P33/41 (annexin IV).
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Highly polarized expression of carbohydrate-binding Protein P33/41 (annexin IV) on the apical plasma membrane of epithelial cells in renal proximal tubules.
FEBS letters, 1994Co-Authors: Kyoko Kojima, Haruko Ogawa, Hideko Utsumi, Isamu MatsumotoAbstract:P33/41 is a Ca2+-dependent carbohydrate-binding Protein and is identical to annexin IV, a member of the annexin Protein family. The localization of P33/41 in bovine kidney specimens was investigated immununohistochemically by use of specific polyclonal antibodies. The most interesting finding on immunostaining was that P33/41 was highly concentrated in the apical plasma membrane of the epithelial cells in the proximal tubules contrary to the distribution throughout the cytoplasm in the papillary ducts and papilla epithelium. The enrichment of P33/41 m the apical membrane was confirmed by immunoblotting of the brush border membrane fraction prepared from a kidney homogenate. Sequential extraction with EDTA and Triton X-100, and a partition experiment with Triton X-114 revealed that most P33/41 associates with the renal brush border membrane in a Ca2+-independent manner and is integrated into the membrane like intrinsic membrane Proteins.
Isamu Matsumoto - One of the best experts on this subject based on the ideXlab platform.
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Characterization of Carbohydrate-binding Protein P33/41: RELATION WITH ANNEXIN IV, MOLECULAR BASIS OF THE DOUBLET FORMS (P33 AND p41), AND MODULATION OF THE CARBOHYDRATE BINDING ACTIVITY BY PHOSPHOLIPIDS (∗)
The Journal of biological chemistry, 1996Co-Authors: Kyoko Kojima, Kazuo Yamamoto, Tatsuro Irimura, Toshiaki Osawa, Haruko Ogawa, Isamu MatsumotoAbstract:A Protein, P33/41, expressed in bovine kidney and many other tissues was identified as a lectin which binds to sialoglycoProteins and glycosaminoglycans in a calcium-dependent manner. Partial amino acid sequences of P33/41 are highly homologous to those of a calcium/phospholipid-binding annexin Protein, annexin IV (endonexin). P33/41 exhibited similar calcium-dependent phospholipid binding activity (Kojima, K., Ogawa, H., Seno, N., Yamamoto, K., Irimura, T., Osawa, T., and Matsumoto, I. (1992) J. Biol. Chem. 267, 20536-20539). To further characterize P33/41, we cloned the P33/41 cDNA and characterized the recombinant Protein encoded by this cDNA. Oligonucleotide probes were synthesized based on partial amino acid sequences of P33/41 and used for screening. A P33/41 cDNA clone was isolated encoding a Protein of 319 amino acids with a calculated molecular mass of 35,769 Da. The deduced amino acid sequence was identical to that of bovine annexin IV except for one amino acid substitution. The recombinant Protein gave two 33-kDa (P33) and 41-kDa (p41) bands on SDS-polyacrylamide gel electrophoresis under non-reducing conditions, and only one 33-kDa band under reducing conditions, as did the native Protein. Mass spectrometric analysis combined with site-directed mutagenesis of each of the four cysteine residues of the recombinant Protein revealed that p41 is a dimer of P33 cross-linked at Cys-198 via a disulfide bond. The recombinant Protein bound to columns of heparin and fetuin glycopeptides in a calcium dependent manner and to phospholipid vesicles composed of phosphatidylserine (PS)/phosphatidylcholine (PC), phosphatidylethanolamine (PE)/PC or phosphatidylinositol (PI)/PC. Furthermore, concurrent binding assays showed that the binding of the recombinant Protein to phospholipid vesicles was not affected by heparin, whereas that to heparin was influenced by the phospholipid composition of the vesicles; the highest binding was observed with vesicles composed of PE/PC. These results suggest that P33/41 binds two types of ligands via different sites and that phospholipids modulate the carbohydrate binding activity of P33/41.
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characterization of carbohydrate binding Protein P33 41 relation with annexin iv molecular basis of the doublet forms P33 and p41 and modulation of the carbohydrate binding activity by phospholipids
Journal of Biological Chemistry, 1996Co-Authors: Kyoko Kojima, Kazuo Yamamoto, Tatsuro Irimura, Toshiaki Osawa, Haruko Ogawa, Isamu MatsumotoAbstract:A Protein, P33/41, expressed in bovine kidney and many other tissues was identified as a lectin which binds to sialoglycoProteins and glycosaminoglycans in a calcium-dependent manner. Partial amino acid sequences of P33/41 are highly homologous to those of a calcium/phospholipid-binding annexin Protein, annexin IV (endonexin). P33/41 exhibited similar calcium-dependent phospholipid binding activity (Kojima, K., Ogawa, H., Seno, N., Yamamoto, K., Irimura, T., Osawa, T., and Matsumoto, I. (1992) J. Biol. Chem. 267, 20536-20539). To further characterize P33/41, we cloned the P33/41 cDNA and characterized the recombinant Protein encoded by this cDNA. Oligonucleotide probes were synthesized based on partial amino acid sequences of P33/41 and used for screening. A P33/41 cDNA clone was isolated encoding a Protein of 319 amino acids with a calculated molecular mass of 35,769 Da. The deduced amino acid sequence was identical to that of bovine annexin IV except for one amino acid substitution. The recombinant Protein gave two 33-kDa (P33) and 41-kDa (p41) bands on SDS-polyacrylamide gel electrophoresis under non-reducing conditions, and only one 33-kDa band under reducing conditions, as did the native Protein. Mass spectrometric analysis combined with site-directed mutagenesis of each of the four cysteine residues of the recombinant Protein revealed that p41 is a dimer of P33 cross-linked at Cys-198 via a disulfide bond. The recombinant Protein bound to columns of heparin and fetuin glycopeptides in a calcium dependent manner and to phospholipid vesicles composed of phosphatidylserine (PS)/phosphatidylcholine (PC), phosphatidylethanolamine (PE)/PC or phosphatidylinositol (PI)/PC. Furthermore, concurrent binding assays showed that the binding of the recombinant Protein to phospholipid vesicles was not affected by heparin, whereas that to heparin was influenced by the phospholipid composition of the vesicles; the highest binding was observed with vesicles composed of PE/PC. These results suggest that P33/41 binds two types of ligands via different sites and that phospholipids modulate the carbohydrate binding activity of P33/41.
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Highly polarized expression of carbohydrate-binding Protein P33/41 (annexin IV) on the apical plasma membrane of epithelial cells in renal proximal tubules.
FEBS letters, 1994Co-Authors: Kyoko Kojima, Haruko Ogawa, Hideko Utsumi, Isamu MatsumotoAbstract:P33/41 is a Ca2+-dependent carbohydrate-binding Protein and is identical to annexin IV, a member of the annexin Protein family. The localization of P33/41 in bovine kidney specimens was investigated immununohistochemically by use of specific polyclonal antibodies. The most interesting finding on immunostaining was that P33/41 was highly concentrated in the apical plasma membrane of the epithelial cells in the proximal tubules contrary to the distribution throughout the cytoplasm in the papillary ducts and papilla epithelium. The enrichment of P33/41 m the apical membrane was confirmed by immunoblotting of the brush border membrane fraction prepared from a kidney homogenate. Sequential extraction with EDTA and Triton X-100, and a partition experiment with Triton X-114 revealed that most P33/41 associates with the renal brush border membrane in a Ca2+-independent manner and is integrated into the membrane like intrinsic membrane Proteins.
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highly polarized expression of carbohydrate binding Protein P33 41 annexin iv on the apical plasma membrane of epithelial cells in renal proximal tubules
FEBS Letters, 1994Co-Authors: Kyoko Kojima, Haruko Ogawa, Hideko Utsumi, Isamu MatsumotoAbstract:P33/41 is a Ca2+-dependent carbohydrate-binding Protein and is identical to annexin IV, a member of the annexin Protein family. The localization of P33/41 in bovine kidney specimens was investigated immununohistochemically by use of specific polyclonal antibodies. The most interesting finding on immunostaining was that P33/41 was highly concentrated in the apical plasma membrane of the epithelial cells in the proximal tubules contrary to the distribution throughout the cytoplasm in the papillary ducts and papilla epithelium. The enrichment of P33/41 m the apical membrane was confirmed by immunoblotting of the brush border membrane fraction prepared from a kidney homogenate. Sequential extraction with EDTA and Triton X-100, and a partition experiment with Triton X-114 revealed that most P33/41 associates with the renal brush border membrane in a Ca2+-independent manner and is integrated into the membrane like intrinsic membrane Proteins.
Svetlana Y. Folimonova - One of the best experts on this subject based on the ideXlab platform.
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Self-interaction of Citrus tristeza virus P33 Protein via N-terminal helix.
Virus research, 2017Co-Authors: Sung-hwan Kang, Thi Nguyet Minh Dao, Ok-kyung Kim, Svetlana Y. FolimonovaAbstract:Citrus tristeza virus (CTV), the most economically important viral pathogen of citrus, encodes a unique Protein, P33. CTV P33 shows no similarity with other known Proteins, yet plays an important role in viral pathogenesis: it extends the virus host range and mediates virus ability to exclude superinfection by other variants of the virus. Previously we demonstrated that P33 is an integral membrane Protein and appears to share characteristics of viral movement Proteins. In this study, we show that the P33 Protein self-interacts in vitro and in vivo using co-immunoprecipitation, yeast two hybrid, and bimolecular fluorescence complementation assays. Furthermore, a helix located at the N-terminus of the Protein is required and sufficient for the Protein self-interaction.
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The conundrum of a unique Protein encoded by citrus tristeza virus that is dispensable for infection of most hosts yet shows characteristics of a viral movement Protein.
Virology, 2015Co-Authors: Aurélie Bak, Svetlana Y. FolimonovaAbstract:Citrus tristeza virus (CTV), one of the most economically important viruses, produces a unique Protein, P33, which is encoded only in the genomes of isolates of CTV. Recently, we demonstrated that membrane association of the P33 Protein confers virus ability to extend its host range. In this work we show that P33 shares characteristics of viral movement Proteins. Upon expression in a host cell, the Protein localizes to plasmodesmata and displays the ability to form extracellular tubules. Furthermore, P33 appears to traffic via the cellular secretory pathway and the actin network to plasmodesmata locations and is likely being recycled through the endocytic pathway. Finally, our study reveals that P33 colocalizes with a putative movement Protein of CTV, the p6 Protein. These results suggest a potential role of P33 as a noncanonical viral movement Protein, which mediates virus translocation in the specific hosts.
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Membrane association of a nonconserved viral Protein confers virus ability to extend its host range.
Virology, 2015Co-Authors: Sung-hwan Kang, Ok-kyung Kim, Aurélie Bak, Svetlana Y. FolimonovaAbstract:Citrus tristeza virus (CTV), the largest and most complex member of the family Closteroviridae, encodes a unique Protein, P33, which shows no homology with other known Proteins, however, plays an important role in virus pathogenesis. In this study, we examined some of the characteristics of P33. We show that P33 is a membrane-associated Protein that is inserted into the membrane via a transmembrane helix formed by hydrophobic amino acid residues at the C-terminal end of the Protein. Removal of this transmembrane domain (TMD) dramatically altered the intracellular localization of P33. Moreover, the TMD alone was sufficient to confer membrane localization of an unrelated Protein. Finally, a CTV variant that produced a truncated P33 lacking the TMD was unable to infect sour orange, one of the selected virus hosts, which infection requires P33, suggesting that membrane association of P33 is important for the ability of CTV to extend its host range.
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Superinfection exclusion by Citrus tristeza virus does not correlate with the production of viral small RNAs.
Virology, 2014Co-Authors: Svetlana Y. Folimonova, Scott J Harper, Michael T Leonard, Eric W Triplett, Turksen ShiltsAbstract:Superinfection exclusion (SIE), a phenomenon in which a preexisting viral infection prevents a secondary infection with the same or closely related virus, has been described for different viruses, including important pathogens of humans, animals, and plants. Several mechanisms acting at various stages of the viral life cycle have been proposed to explain SIE. Most cases of SIE in plant virus systems were attributed to induction of RNA silencing, a host defense mechanism that is mediated by small RNAs. Here we show that SIE by Citrus tristeza virus (CTV) does not correlate with the production of viral small interfering RNAs (siRNAs). CTV variants, which differed in the SIE ability, had similar siRNAs profiles. Along with our previous observations that the exclusion phenomenon requires a specific viral Protein, P33, the new data suggest that SIE by CTV is highly complex and appears to use different mechanisms than those proposed for other viruses.
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A Viral Protein Mediates Superinfection Exclusion at the Whole-Organism Level but Is Not Required for Exclusion at the Cellular Level
Journal of virology, 2014Co-Authors: María Bergua, Turksen Shilts, Mark P. Zwart, Choaa El-mohtar, Santiago F. Elena, Svetlana Y. FolimonovaAbstract:Superinfection exclusion (SIE), the ability of an established virus infection to interfere with a secondary infection by the same or a closely related virus, has been described for different viruses, including important pathogens of humans, animals, and plants. Citrus tristeza virus (CTV), a positive-sense RNA virus, represents a valuable model system for studying SIE due to the existence of several phylogenetically distinct strains. Furthermore, CTV allows SIE to be examined at the whole-organism level. Previously, we demonstrated that SIE by CTV is a virus-controlled function that requires the viral Protein P33. In this study, we show that P33 mediates SIE at the whole-organism level, while it is not required for exclusion at the cellular level. Primary infection of a host with a fluorescent Protein-tagged CTV variant lacking P33 did not interfere with the establishment of a secondary infection by the same virus labeled with a different fluorescent Protein. However, cellular coinfection by both viruses was rare. The obtained observations, along with estimates of the cellular multiplicity of infection (MOI) and MOI model selection, suggested that low levels of cellular coinfection appear to be best explained by exclusion at the cellular level. Based on these results, we propose that SIE by CTV is operated at two levels—the cellular and the whole-organism levels—by two distinct mechanisms that could function independently. This novel aspect of viral SIE highlights the intriguing complexity of this phenomenon, further understanding of which may open up new avenues to manage virus diseases. IMPORTANCE Many viruses exhibit superinfection exclusion (SIE), the ability of an established virus infection to interfere with a secondary infection by related viruses. SIE plays an important role in the pathogenesis and evolution of virus populations. The observations described here suggest that SIE could be controlled independently at different levels of the host: the whole-organism level or the level of individual cells. The P33 Protein of citrus tristeza virus (CTV), an RNA virus, was shown to mediate SIE at the whole-organism level, while it appeared not to be required for exclusion at the cellular level. SIE by CTV is, therefore, highly complex and appears to use mechanisms different from those proposed for other viruses. A better understanding of this phenomenon may lead to the development of new strategies for controlling viral diseases in human populations and agroecosystems.
Haruko Ogawa - One of the best experts on this subject based on the ideXlab platform.
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Characterization of Carbohydrate-binding Protein P33/41: RELATION WITH ANNEXIN IV, MOLECULAR BASIS OF THE DOUBLET FORMS (P33 AND p41), AND MODULATION OF THE CARBOHYDRATE BINDING ACTIVITY BY PHOSPHOLIPIDS (∗)
The Journal of biological chemistry, 1996Co-Authors: Kyoko Kojima, Kazuo Yamamoto, Tatsuro Irimura, Toshiaki Osawa, Haruko Ogawa, Isamu MatsumotoAbstract:A Protein, P33/41, expressed in bovine kidney and many other tissues was identified as a lectin which binds to sialoglycoProteins and glycosaminoglycans in a calcium-dependent manner. Partial amino acid sequences of P33/41 are highly homologous to those of a calcium/phospholipid-binding annexin Protein, annexin IV (endonexin). P33/41 exhibited similar calcium-dependent phospholipid binding activity (Kojima, K., Ogawa, H., Seno, N., Yamamoto, K., Irimura, T., Osawa, T., and Matsumoto, I. (1992) J. Biol. Chem. 267, 20536-20539). To further characterize P33/41, we cloned the P33/41 cDNA and characterized the recombinant Protein encoded by this cDNA. Oligonucleotide probes were synthesized based on partial amino acid sequences of P33/41 and used for screening. A P33/41 cDNA clone was isolated encoding a Protein of 319 amino acids with a calculated molecular mass of 35,769 Da. The deduced amino acid sequence was identical to that of bovine annexin IV except for one amino acid substitution. The recombinant Protein gave two 33-kDa (P33) and 41-kDa (p41) bands on SDS-polyacrylamide gel electrophoresis under non-reducing conditions, and only one 33-kDa band under reducing conditions, as did the native Protein. Mass spectrometric analysis combined with site-directed mutagenesis of each of the four cysteine residues of the recombinant Protein revealed that p41 is a dimer of P33 cross-linked at Cys-198 via a disulfide bond. The recombinant Protein bound to columns of heparin and fetuin glycopeptides in a calcium dependent manner and to phospholipid vesicles composed of phosphatidylserine (PS)/phosphatidylcholine (PC), phosphatidylethanolamine (PE)/PC or phosphatidylinositol (PI)/PC. Furthermore, concurrent binding assays showed that the binding of the recombinant Protein to phospholipid vesicles was not affected by heparin, whereas that to heparin was influenced by the phospholipid composition of the vesicles; the highest binding was observed with vesicles composed of PE/PC. These results suggest that P33/41 binds two types of ligands via different sites and that phospholipids modulate the carbohydrate binding activity of P33/41.
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characterization of carbohydrate binding Protein P33 41 relation with annexin iv molecular basis of the doublet forms P33 and p41 and modulation of the carbohydrate binding activity by phospholipids
Journal of Biological Chemistry, 1996Co-Authors: Kyoko Kojima, Kazuo Yamamoto, Tatsuro Irimura, Toshiaki Osawa, Haruko Ogawa, Isamu MatsumotoAbstract:A Protein, P33/41, expressed in bovine kidney and many other tissues was identified as a lectin which binds to sialoglycoProteins and glycosaminoglycans in a calcium-dependent manner. Partial amino acid sequences of P33/41 are highly homologous to those of a calcium/phospholipid-binding annexin Protein, annexin IV (endonexin). P33/41 exhibited similar calcium-dependent phospholipid binding activity (Kojima, K., Ogawa, H., Seno, N., Yamamoto, K., Irimura, T., Osawa, T., and Matsumoto, I. (1992) J. Biol. Chem. 267, 20536-20539). To further characterize P33/41, we cloned the P33/41 cDNA and characterized the recombinant Protein encoded by this cDNA. Oligonucleotide probes were synthesized based on partial amino acid sequences of P33/41 and used for screening. A P33/41 cDNA clone was isolated encoding a Protein of 319 amino acids with a calculated molecular mass of 35,769 Da. The deduced amino acid sequence was identical to that of bovine annexin IV except for one amino acid substitution. The recombinant Protein gave two 33-kDa (P33) and 41-kDa (p41) bands on SDS-polyacrylamide gel electrophoresis under non-reducing conditions, and only one 33-kDa band under reducing conditions, as did the native Protein. Mass spectrometric analysis combined with site-directed mutagenesis of each of the four cysteine residues of the recombinant Protein revealed that p41 is a dimer of P33 cross-linked at Cys-198 via a disulfide bond. The recombinant Protein bound to columns of heparin and fetuin glycopeptides in a calcium dependent manner and to phospholipid vesicles composed of phosphatidylserine (PS)/phosphatidylcholine (PC), phosphatidylethanolamine (PE)/PC or phosphatidylinositol (PI)/PC. Furthermore, concurrent binding assays showed that the binding of the recombinant Protein to phospholipid vesicles was not affected by heparin, whereas that to heparin was influenced by the phospholipid composition of the vesicles; the highest binding was observed with vesicles composed of PE/PC. These results suggest that P33/41 binds two types of ligands via different sites and that phospholipids modulate the carbohydrate binding activity of P33/41.
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Expression of Carbohydrate-Binding Protein P33/41 in Human Tumor Cell Lines
Journal of biochemistry, 1996Co-Authors: Ayano Satoh, Kyoko Kojima, Haruko Ogawa, Eiji Takayama, Takao Yamori, Shigeo Sato, Tokuichi Kawaguchi, Takashi Tsuruo, Yoshimoto Katsura, Tatsuo KinaAbstract:We previously reported a new type of lectin, P33/41 (annexin IV), which was isolated from a bovine tissue extract [Kojima, K. et al. (1992) J. Biol. Chem. 267, 20536-20539]. When the expression of P33/41 (annexin IV) was surveyed in the lysates of 39 human tumor cell lines by SDS-PAGE, followed by Western blot analysis with polyclonal anti-bovine P33/41 and monoclonal anti-annexin IV (Z016, Zymed) antibodies, 21 cell lines were found to be reactive with the polyclonal antibody, whereas all 39 cell lines were stained with Z016. These results together with those obtained with standard Proteins, annexins IV and V, suggested that the monoclonal antibody, Z016, recognizes annexin V, but not P33/41 (annexin IV). Therefore, we performed cDNA cloning of human P33/41 (annexin IV) to prepare a recombinant Protein and raised monoclonal antibodies against the Protein. Northern blot analysis with the cDNA as a probe showed that a human colon cancer cell line, HT29, contains P33/41 (annexin IV) mRNA of two sizes, 2.0 and 3.0 kb. The two monoclonal antibodies, AS11 and AS17, against the recombinant Protein generated were useful for flow cytometric analysis, ELISA, Western blot analysis and immunoprecipitation. Flow cytometric analysis with AS17 showed that P33/41 (annexin IV) is located in the cytoplasm of HT29 cells, but not on the cell surface. However, one of the cell surface Proteins first labeled with biotin and then solubilized with a detergent was immunoprecipitated with AS17. The results suggest the existence of a membrane spanning form of P33/41 (annexin IV).
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expression of carbohydrate binding Protein P33 41 in human tumor cell lines
Journal of Biochemistry, 1996Co-Authors: Ayano Satoh, Kyoko Kojima, Haruko Ogawa, Eiji Takayama, Takao Yamori, Shigeo Sato, Tokuichi Kawaguchi, Takashi Tsuruo, Yoshimoto Katsura, Tatsuo KinaAbstract:We previously reported a new type of lectin, P33/41 (annexin IV), which was isolated from a bovine tissue extract [Kojima, K. et al. (1992) J. Biol. Chem. 267, 20536-20539]. When the expression of P33/41 (annexin IV) was surveyed in the lysates of 39 human tumor cell lines by SDS-PAGE, followed by Western blot analysis with polyclonal anti-bovine P33/41 and monoclonal anti-annexin IV (Z016, Zymed) antibodies, 21 cell lines were found to be reactive with the polyclonal antibody, whereas all 39 cell lines were stained with Z016. These results together with those obtained with standard Proteins, annexins IV and V, suggested that the monoclonal antibody, Z016, recognizes annexin V, but not P33/41 (annexin IV). Therefore, we performed cDNA cloning of human P33/41 (annexin IV) to prepare a recombinant Protein and raised monoclonal antibodies against the Protein. Northern blot analysis with the cDNA as a probe showed that a human colon cancer cell line, HT29, contains P33/41 (annexin IV) mRNA of two sizes, 2.0 and 3.0 kb. The two monoclonal antibodies, AS11 and AS17, against the recombinant Protein generated were useful for flow cytometric analysis, ELISA, Western blot analysis and immunoprecipitation. Flow cytometric analysis with AS17 showed that P33/41 (annexin IV) is located in the cytoplasm of HT29 cells, but not on the cell surface. However, one of the cell surface Proteins first labeled with biotin and then solubilized with a detergent was immunoprecipitated with AS17. The results suggest the existence of a membrane spanning form of P33/41 (annexin IV).
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Highly polarized expression of carbohydrate-binding Protein P33/41 (annexin IV) on the apical plasma membrane of epithelial cells in renal proximal tubules.
FEBS letters, 1994Co-Authors: Kyoko Kojima, Haruko Ogawa, Hideko Utsumi, Isamu MatsumotoAbstract:P33/41 is a Ca2+-dependent carbohydrate-binding Protein and is identical to annexin IV, a member of the annexin Protein family. The localization of P33/41 in bovine kidney specimens was investigated immununohistochemically by use of specific polyclonal antibodies. The most interesting finding on immunostaining was that P33/41 was highly concentrated in the apical plasma membrane of the epithelial cells in the proximal tubules contrary to the distribution throughout the cytoplasm in the papillary ducts and papilla epithelium. The enrichment of P33/41 m the apical membrane was confirmed by immunoblotting of the brush border membrane fraction prepared from a kidney homogenate. Sequential extraction with EDTA and Triton X-100, and a partition experiment with Triton X-114 revealed that most P33/41 associates with the renal brush border membrane in a Ca2+-independent manner and is integrated into the membrane like intrinsic membrane Proteins.
