The Experts below are selected from a list of 102 Experts worldwide ranked by ideXlab platform
Hiroyoshi Hidaka - One of the best experts on this subject based on the ideXlab platform.
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S100 beta is a target protein of neurocalcin delta, an abundant isoform in glial cells.
Biochemical Journal, 1995Co-Authors: K. Okazaki, N H Obata, S. Inoue, Hiroyoshi HidakaAbstract:To clarify the function of neurocalcin delta, an isoform found abundantly in glial cells, we attempted to find its target proteins by using neurocalcin delta-affinity chromatography and the 125I-neurocalcin delta gel-overlay method. The 10, 14, 27, 36 and 50 kDa bands found on SDS/PAGE bound to 125I-neurocalcin delta, and 10, 11, 19, 24, 26, 50 and 70 kDa proteins were eluted from a neurocalcin delta-affinity column in a Ca(2+)-dependent manner. Sequence analysis of proteolytic peptides revealed the following identities: S100 beta (10 kDa), S100 alpha (11 kDa), myelin basic protein (19 kDa), glyceraldehyde-3-phosphate dehydrogenase (36 kDa) and tubulin beta-chain (50 kDa). A zero-length cross-linking study indicated that 1 mol of S100 beta bound to 1 mol of neurocalcin delta. With the gel-overlay method, purified S100 beta protein and calcyclin bound to 125I-neurocalcin delta whereas calgizarrin and Calvasculin, other members of the S100 family, did not. These findings suggest that S100 beta is one of the target proteins of neurocalcin delta, and the neurocalcin delta-S100 beta complex may be involved in Ca(2+)-signalling in the glial cell.
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Calvasculin, as a factor affecting the microfilament assemblies in rat fibroblasts transfected by src gene
FEBS letters, 1993Co-Authors: Yasuo Watanabe, Nobuteru Usada, Hiroyuki Minami, Takashi Morita, Shinichiro Tsugane, Ryoki Ishikawa, Kazuhiro Kohama, Yasuhiro Tomida, Hiroyoshi HidakaAbstract:Cell transformations accompany alterations in cell morphology and microfilament patterns. Calvasculin encodes mRNA termed pEL-98, 18A2, 42A, p9Ka, or mtsl, found to be elevated in several metastatic cell lines. We report the elevation of Calvasculin expression in SR-3Y1 cells, which show disappearance of ordered microfilaments, compared to that in 3Y1 cells and that the similar distribution of Calvasculin to that of actin filaments. Interestingly, Calvasculin co-sediments with F-actin and bundles actin filaments in a Ca2+-dependent manner. This activity, along with the elevation of Calvasculin following transformation, suggests that the disorganization of filaments in SR-3Y1 cell is due to the cross-linking activity of Calvasculin.
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Calcyclin and Calvasculin exist in human platelets.
Biochemical and biophysical research communications, 1992Co-Authors: Yasuhiro Tomida, Ryoji Kobayashi, Motomu Terasawa, Hiroyoshi HidakaAbstract:Abstract Using Ca2+-dependent affinity chromatography on a synthetic compound (W-7)-coupled Sepharose column, three distinct Ca2+-binding proteins have been identified in human platelets. The molecular mass of these three distinct proteins was estimated to be 10, 10.5, 17 kDa, respectively, by polyacrylamide gel electrophoresis in the presence of SDS. The partial amino acid sequence revealed these proteins have EF-hand structures and high homology to the predicted proteins, calcyclin, Calvasculin, and calmodulin. Calcyclin and Calvasculin have been considered as probably having roles in the control of cell proliferation, but the existence of these two proteins in platelets suggests that they have other intracellular functions related to the Ca2+-signal transduction system.
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Calvasculin, an encoded protein from mRNA termed pEL-98, 18A2, 42A, or p9Ka, is secreted by smooth muscle cells in culture and exhibits Ca(2+)-dependent binding to 36-kDa microfibril-associated glycoprotein.
The Journal of biological chemistry, 1992Co-Authors: Yoshihito Watanabe, Ryoji Kobayashi, N Usuda, S Tsugane, Hiroyoshi HidakaAbstract:Abstract Calvasculin, an EF-hand protein with a molecular mass of 11 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, is present abundantly in bovine aorta (Watanabe, Y., Kobayashi, R., Ishikawa, T., and Hidaka, H. (1992) Arch. Biochem. Biophys. 292, 563-569). This protein is synthesized constitutively by bovine aortic smooth muscle (BASM) cells and rat embryo fibroblast 3Y1 cells in culture. We discovered that Calvasculin was secreted by BASM cells and 3Y1 cells. Immunofluorescence staining of BASM cells showed a granular distribution for Calvasculin that was typical of a secreted protein. This protein bound with an extracellular matrix protein, 36-kDa microfibril-associated glycoprotein (36-kDa MAP), in a Ca(2+)-dependent manner in vitro. A stoichiometry analysis showed that the 36-kDa MAP bound 2.2 Calvasculin eq/mol of protein. Solid-phase binding assays indicated a preferential affinity of native Calvasculin for 36-kDa MAP among the extracellular matrices in a Ca(2+)-dependent manner. These results suggest that Calvasculin, intracellular Ca(2+)-binding protein, is released to the extracellular space and binds with 36-kDa MAP.
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Specific binding of CAP-50 to calcyclin.
FEBS letters, 1992Co-Authors: Hiroyuki Minami, Yasuo Watanabe, Hiroshi Tokumitsu, Akihiro Mizutani, Masato Watanabe, Hiroyoshi HidakaAbstract:CAP-50, a calcyclin-associated protein with an apparent molecular mass or 50 kDa, was purified and proved to be a novel annexin [Tokumitsu, H. et al. (1992) J. Biol. Chem. 267, 8919–8924]. We examined the binding of CAP-50 to other Ca2+-binding proteins which have two or four EF-hand structures, by a co-precipitation assay with phospholipid (phosphatidylserine). Among nine Ca2+-binding proteins (calcyclin, S-100 proteins, p11, calgizzarin, Calvasculin, calmodulin and troponin C) examined, only calcyclin interacted with CAP-50. These results clearly show that the interaction of CAP-50 to calcyclin is specific, i.e. other Ca2+-binding proteins with the EF-hand structure could not substitute for calcyclin, thereby suggesting the possible role in specific regulation of the function of CAP-50 by Ca2+/calcyclin.
Roger Barraclough - One of the best experts on this subject based on the ideXlab platform.
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Expression of the rat, S-100-related, calcium-binding protein gene, p9Ka, in transgenic mice demonstrates different patterns of expression between these two species.
DNA and cell biology, 1995Co-Authors: Michael P.a. Davies, Stephen E. Harris, Philip S. Rudland, Roger BarracloughAbstract:ABSTRACT p9Ka (also known as mts 1/18A2/Calvasculin/CAPL) is a member of the S-100-related family of small, calcium-binding proteins. Previous studies suggest apparent discrepancies between the exp...
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interactions in vitro of p9ka the rat s 100 related metastasis inducing calcium binding protein
Journal of Biological Chemistry, 1994Co-Authors: F E M Gibbs, Philip S. Rudland, M Wilkinson, Roger BarracloughAbstract:Abstract The S-100 proteins are a structurally related family displaying diverse intracellular and extracellular interactions. One such protein p9Ka (also known as Calvasculin), or its mRNA (also known as CAPL, 42A, 18A2, mts, pEL 98), becomes elevated upon changes in the growth and differentiation of cells. Overexpression of p9Ka in benign rat mammary cells induces the metastatic phenotype. In order to help understand the role of p9Ka in these processes, the molecular properties of recombinant rat p9Ka have been studied. Recombinant p9Ka forms multimers in vitro, which are not due to intermolecular disulfide bridges, it binds 2 mol of calcium ions/mol of protein, and the binding of calcium ions is strongly antagonized by monovalent and divalent cations tested. Immunofluorescence studies indicate that p9Ka is located on cytoskeletal elements in a pattern which is identical to actin filaments stained with phalloidin. In vitro, it is shown that recombinant p9Ka binds to sites on at least two intracellular polypeptides. These sites display the same binding capacity for p9Ka in extracts of cultured rat mammary cells which show widely differing levels of expression of natural p9Ka. The results suggest that the production of p9Ka, and not of its target molecules, may be associated with the changes seen in cultured cells.
Yasuo Watanabe - One of the best experts on this subject based on the ideXlab platform.
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Calvasculin, as a factor affecting the microfilament assemblies in rat fibroblasts transfected by src gene
FEBS letters, 1993Co-Authors: Yasuo Watanabe, Nobuteru Usada, Hiroyuki Minami, Takashi Morita, Shinichiro Tsugane, Ryoki Ishikawa, Kazuhiro Kohama, Yasuhiro Tomida, Hiroyoshi HidakaAbstract:Cell transformations accompany alterations in cell morphology and microfilament patterns. Calvasculin encodes mRNA termed pEL-98, 18A2, 42A, p9Ka, or mtsl, found to be elevated in several metastatic cell lines. We report the elevation of Calvasculin expression in SR-3Y1 cells, which show disappearance of ordered microfilaments, compared to that in 3Y1 cells and that the similar distribution of Calvasculin to that of actin filaments. Interestingly, Calvasculin co-sediments with F-actin and bundles actin filaments in a Ca2+-dependent manner. This activity, along with the elevation of Calvasculin following transformation, suggests that the disorganization of filaments in SR-3Y1 cell is due to the cross-linking activity of Calvasculin.
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Specific binding of CAP-50 to calcyclin.
FEBS letters, 1992Co-Authors: Hiroyuki Minami, Yasuo Watanabe, Hiroshi Tokumitsu, Akihiro Mizutani, Masato Watanabe, Hiroyoshi HidakaAbstract:CAP-50, a calcyclin-associated protein with an apparent molecular mass or 50 kDa, was purified and proved to be a novel annexin [Tokumitsu, H. et al. (1992) J. Biol. Chem. 267, 8919–8924]. We examined the binding of CAP-50 to other Ca2+-binding proteins which have two or four EF-hand structures, by a co-precipitation assay with phospholipid (phosphatidylserine). Among nine Ca2+-binding proteins (calcyclin, S-100 proteins, p11, calgizzarin, Calvasculin, calmodulin and troponin C) examined, only calcyclin interacted with CAP-50. These results clearly show that the interaction of CAP-50 to calcyclin is specific, i.e. other Ca2+-binding proteins with the EF-hand structure could not substitute for calcyclin, thereby suggesting the possible role in specific regulation of the function of CAP-50 by Ca2+/calcyclin.
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Isolation and characterization of a calcium-binding protein derived from mRNA termed p9Ka, pEL-98, 18A2, or 42A by the newly synthesized vasorelaxant W-66 affinity chromatography.
Archives of biochemistry and biophysics, 1992Co-Authors: Yasuo Watanabe, Ryoji Kobayashi, Tomohiko Ishikawa, Hiroyoshi HidakaAbstract:W-66 (N-(2-aminoethyl)-N-[2-(4-chlorocinnamylamino) ethyl]-5-isoquinolinesulfonamide), a newly synthesized isoquinolinesulfonamide, was shown to have a potent vasodilatory action and calmodulin (CaM)-antagonizing action. Using the W-66 affinity chromatographic technique, we purified two Ca2+-binding proteins from the EGTA-soluble fraction of bovine aorta. One was CaM and the other was an acidic protein with a molecular mass of 11 kDa. It was tentatively named “Calvasculin”. Calvasculin was a dimeric protein. Equilibrium dialysis showed that 1 mol of Calvasculin (dimer) bound to 1.98 ± 0.30 mol of Ca2+ in the presence of 10−3m Ca2+. Calvasculin failed to activate Ca2+CaM-dependent enzymes such as myosin light chain kinase, Ca2+CaM-dependent phosphodiesterase, or Ca2+CaM-dependent protein kinase II and to inhibit the CaM stimulation of these enzymes. The partial amino acid sequence of Calvasculin revealed a high homology to the predicted protein derived from mRNA, named pEL-98, 18A2, 42A, or p9Ka. We also examined the physicochemical and biochemical properties of Calvasculin. Using the antibody specific for Calvasculin, we obtained evidence that Calvasculin is present in abundance in bovine aorta but not in brain, lung, heart, or testis.
Ryoji Kobayashi - One of the best experts on this subject based on the ideXlab platform.
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Calcyclin and Calvasculin exist in human platelets.
Biochemical and biophysical research communications, 1992Co-Authors: Yasuhiro Tomida, Ryoji Kobayashi, Motomu Terasawa, Hiroyoshi HidakaAbstract:Abstract Using Ca2+-dependent affinity chromatography on a synthetic compound (W-7)-coupled Sepharose column, three distinct Ca2+-binding proteins have been identified in human platelets. The molecular mass of these three distinct proteins was estimated to be 10, 10.5, 17 kDa, respectively, by polyacrylamide gel electrophoresis in the presence of SDS. The partial amino acid sequence revealed these proteins have EF-hand structures and high homology to the predicted proteins, calcyclin, Calvasculin, and calmodulin. Calcyclin and Calvasculin have been considered as probably having roles in the control of cell proliferation, but the existence of these two proteins in platelets suggests that they have other intracellular functions related to the Ca2+-signal transduction system.
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Calvasculin, an encoded protein from mRNA termed pEL-98, 18A2, 42A, or p9Ka, is secreted by smooth muscle cells in culture and exhibits Ca(2+)-dependent binding to 36-kDa microfibril-associated glycoprotein.
The Journal of biological chemistry, 1992Co-Authors: Yoshihito Watanabe, Ryoji Kobayashi, N Usuda, S Tsugane, Hiroyoshi HidakaAbstract:Abstract Calvasculin, an EF-hand protein with a molecular mass of 11 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, is present abundantly in bovine aorta (Watanabe, Y., Kobayashi, R., Ishikawa, T., and Hidaka, H. (1992) Arch. Biochem. Biophys. 292, 563-569). This protein is synthesized constitutively by bovine aortic smooth muscle (BASM) cells and rat embryo fibroblast 3Y1 cells in culture. We discovered that Calvasculin was secreted by BASM cells and 3Y1 cells. Immunofluorescence staining of BASM cells showed a granular distribution for Calvasculin that was typical of a secreted protein. This protein bound with an extracellular matrix protein, 36-kDa microfibril-associated glycoprotein (36-kDa MAP), in a Ca(2+)-dependent manner in vitro. A stoichiometry analysis showed that the 36-kDa MAP bound 2.2 Calvasculin eq/mol of protein. Solid-phase binding assays indicated a preferential affinity of native Calvasculin for 36-kDa MAP among the extracellular matrices in a Ca(2+)-dependent manner. These results suggest that Calvasculin, intracellular Ca(2+)-binding protein, is released to the extracellular space and binds with 36-kDa MAP.
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Isolation and characterization of a calcium-binding protein derived from mRNA termed p9Ka, pEL-98, 18A2, or 42A by the newly synthesized vasorelaxant W-66 affinity chromatography.
Archives of biochemistry and biophysics, 1992Co-Authors: Yasuo Watanabe, Ryoji Kobayashi, Tomohiko Ishikawa, Hiroyoshi HidakaAbstract:W-66 (N-(2-aminoethyl)-N-[2-(4-chlorocinnamylamino) ethyl]-5-isoquinolinesulfonamide), a newly synthesized isoquinolinesulfonamide, was shown to have a potent vasodilatory action and calmodulin (CaM)-antagonizing action. Using the W-66 affinity chromatographic technique, we purified two Ca2+-binding proteins from the EGTA-soluble fraction of bovine aorta. One was CaM and the other was an acidic protein with a molecular mass of 11 kDa. It was tentatively named “Calvasculin”. Calvasculin was a dimeric protein. Equilibrium dialysis showed that 1 mol of Calvasculin (dimer) bound to 1.98 ± 0.30 mol of Ca2+ in the presence of 10−3m Ca2+. Calvasculin failed to activate Ca2+CaM-dependent enzymes such as myosin light chain kinase, Ca2+CaM-dependent phosphodiesterase, or Ca2+CaM-dependent protein kinase II and to inhibit the CaM stimulation of these enzymes. The partial amino acid sequence of Calvasculin revealed a high homology to the predicted protein derived from mRNA, named pEL-98, 18A2, 42A, or p9Ka. We also examined the physicochemical and biochemical properties of Calvasculin. Using the antibody specific for Calvasculin, we obtained evidence that Calvasculin is present in abundance in bovine aorta but not in brain, lung, heart, or testis.
Keizo Takenaga - One of the best experts on this subject based on the ideXlab platform.
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Increased expression of S100A4, a metastasis-associated gene, in human colorectal adenocarcinomas.
Clinical Cancer Research, 1997Co-Authors: Keizo Takenaga, H Nakanishi, K Wada, M Suzuki, O Matsuzaki, A Matsuura, Hideya EndoAbstract:The S100A4 gene (also known as pEL98/mts1/p9Ka/18A2/42A/Calvasculin /FSP1/CAPL) encoding an S100-related calcium-binding protein is implied to be involved in the invasion and metastasis of murine tumor cells. In the present study, the expression of S100A4 in human colorectal adenocarcinoma cell lines (SW837, LoVo, DLD-1, HT-29, SW480, SW620, WiDr, and Colo201) and surgically resected neoplastic tissues was examined to investigate whether S100A4 plays a role in the invasion and metastasis of human tumor cells. Northern blot analysis using total RNA isolated from the adenocarcinoma cell lines revealed that five of the eight cell lines expressed substantial amounts of S100A4 mRNA. Normal colon fibroblasts (CCD-18Co) expressed little of the RNA. Using surgically resected specimens, it seemed that the amount of S100A4 mRNA in adenomas was nearly equal to that in normal colonic mucosa, whereas adenocarcinomas expressed a significantly higher amount of the RNA than did the adjacent normal colonic mucosa. Immunohistochemical analysis using formalin-fixed paraffin-embedded surgical specimens and monoclonal anti-S100A4 antibody demonstrated that none of 12 adenoma specimens were immunopositive, whereas 8 of 18 (44%) focal carcinomas in carcinoma in adenoma specimens and 50 of 53 (94%) adenocarcinoma specimens were immunopositive. Interestingly, the incidence of immunopositive cells increased according to the depth of invasion, and nearly all of the carcinoma cells in 14 metastases in the liver were positive. These results suggest that S100A4 may be involved in the progression and the metastatic process of human colorectal neoplastic cells.
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Expression of antisense RNA to S100A4 gene encoding an S100-related calcium-binding protein suppresses metastatic potential of high-metastatic Lewis lung carcinoma cells
Oncogene, 1997Co-Authors: Keizo Takenaga, Yohko Nakamura, Shigeru SakiyamaAbstract:S100A4 (also known as pEL98/mts1/p9Ka/18A2/42A/Calvasculin/FSP1/CAPL), a member of S100-related calcium-binding proteins, has been implicated to play a role in metastasis. In the present study, we examined the effect of antisense S100A4 RNA on metastatic potential of Lewis lung carcinoma (LLC) cells. High-metastatic A11 cells were transfected with the expression vector containing S100A4 cDNA in an inverted (antisense) orientation under the transcriptional control of the mouse metallothionein promoter. Treatment of a stably transfected clone (AS10 cells) with Zn^2+ resulted in the suppression of the experimental metastatic ability, which was accompanied with the expression of antisense S100A4 RNA and the suppression of the S100A4 expression at both the mRNA and the protein levels. To further confirm the effect of antisense S100A4 RNA, we established several clones after retroviral transduction with an antisense S100A4 construct. Notably, reduced metastatic potential was also evident in these clones. In the antisense S100A4 RNA-expressing cells, cell motility and in vitro invasiveness were found to be suppressed.
