The Experts below are selected from a list of 225 Experts worldwide ranked by ideXlab platform
Kazuhiro Yamamoto - One of the best experts on this subject based on the ideXlab platform.
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pericellular proteolysis by matrix metalloproteinase 7 is differentially modulated by Cholesterol Sulfate sulfatide and cardiolipin
FEBS Journal, 2014Co-Authors: Kazuhiro Yamamoto, Kaoru Miyazaki, Shouichi HigashiAbstract:Matrix metalloproteinase (MMP)-7 binds to cell surface Cholesterol Sulfate (CS) and acts as a membrane-associated protease. We have previously found that CS modulates the substrate preference of MMP-7, thereby regulating its pericellular proteolytic action. MMP-7 potentially associates with the cell surface via sulfatide (SM4) and cardiolipin (CL) when they are overexpressed on the cell surface. Here, we investigated the molecular interaction between these acidic lipids and MMP-7 or its substrates, and their effects on the activity of MMP-7. Studies using MMP-7 variants with low CS-binding ability suggested that these lipids interact with a similar site on MMP-7. The hydroxamate-based MMP inhibitor TAPI-1 markedly reduced the affinity of MMP-7 for CS and CL, whereas that for SM4 was not affected by TAPI-1. These three acidic lipids also had different effects on the hydrolytic activity of MMP-7 towards a small peptide substrate: SM4, CL and CS reduced the activity to 80%, 92%, and 20%, respectively. Nevertheless, SM4 and CS similarly accelerated the MMP-7-catalyzed degradation of fibronectin and laminin-332, whereas CL did not. The increased proteolysis of substrate was observed only when both substrate and enzyme had affinity for the lipid, suggesting that the lipids probably bring the reactants into closer proximity. Furthermore, MMP-7 bound to cell surface SM4 or CS cleaved specific cell surface proteins and released similar fragments, whereas the cleavage was not stimulated by cell surface CL-bound MMP-7. This study provides a novel mechanism by which acidic lipids differentially regulate pericellular proteolysis by MMP-7 through allosteric alteration of the substrate-binding site and their inherent affinities for MMP-7 substrates.
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Pericellular proteolysis by matrix metalloproteinase‐7 is differentially modulated by Cholesterol Sulfate, sulfatide, and cardiolipin
The FEBS journal, 2014Co-Authors: Kazuhiro Yamamoto, Kaoru Miyazaki, Shouichi HigashiAbstract:Matrix metalloproteinase (MMP)-7 binds to cell surface Cholesterol Sulfate (CS) and acts as a membrane-associated protease. We have previously found that CS modulates the substrate preference of MMP-7, thereby regulating its pericellular proteolytic action. MMP-7 potentially associates with the cell surface via sulfatide (SM4) and cardiolipin (CL) when they are overexpressed on the cell surface. Here, we investigated the molecular interaction between these acidic lipids and MMP-7 or its substrates, and their effects on the activity of MMP-7. Studies using MMP-7 variants with low CS-binding ability suggested that these lipids interact with a similar site on MMP-7. The hydroxamate-based MMP inhibitor TAPI-1 markedly reduced the affinity of MMP-7 for CS and CL, whereas that for SM4 was not affected by TAPI-1. These three acidic lipids also had different effects on the hydrolytic activity of MMP-7 towards a small peptide substrate: SM4, CL and CS reduced the activity to 80%, 92%, and 20%, respectively. Nevertheless, SM4 and CS similarly accelerated the MMP-7-catalyzed degradation of fibronectin and laminin-332, whereas CL did not. The increased proteolysis of substrate was observed only when both substrate and enzyme had affinity for the lipid, suggesting that the lipids probably bring the reactants into closer proximity. Furthermore, MMP-7 bound to cell surface SM4 or CS cleaved specific cell surface proteins and released similar fragments, whereas the cleavage was not stimulated by cell surface CL-bound MMP-7. This study provides a novel mechanism by which acidic lipids differentially regulate pericellular proteolysis by MMP-7 through allosteric alteration of the substrate-binding site and their inherent affinities for MMP-7 substrates.
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Identification of Amino Acid Residues of Matrix Metalloproteinase-7 Essential for Binding to Cholesterol Sulfate
The Journal of biological chemistry, 2008Co-Authors: Shouichi Higashi, Kazuhiro Yamamoto, Miwa Oeda, Kaoru MiyazakiAbstract:Matrix metalloproteinase-7 (MMP-7; matrilysin) induces homotypic adhesion of colon cancer cells by cleaving cell surface protein(s) and enhances their metastatic potential. Our previous study (Yamamoto, K., Higashi, S., Kioi, M., Tsunezumi, J., Honke, K., and Miyazaki, K. (2006) J. Biol. Chem. 281, 9170-9180) demonstrated that binding of MMP-7 to cell surface Cholesterol Sulfate (CS) is essential for the cell membrane-associated proteolytic action of the protease. To determine the region of MMP-7 essential for binding to CS, we constructed chimeric proteases consisting of various parts of MMP-7 and those of the catalytic domain of MMP-2; the latter protease does not have an affinity for CS. Studies of these chimeric proteases and other mutants of MMP-7 revealed that Ile29, Arg33, Arg51, and Trp55, in the internal sequence, and the C-terminal three residues corresponding to residues 171-173 of MMP-7 are essential for binding to CS. An MMP-7 mutant, which had the internal 4 residues at positions 29, 33, 51, and 55 of MMP-7 replaced with the corresponding residues of MMP-2 and the C-terminal 3 residues deleted, had essentially no affinity for CS. This mutant and wild-type MMP-7 showed similar proteolytic activity toward fibronectin, whereas the mutant lacked the ability to induce the colon cancer cell aggregation. In the three-dimensional structure of MMP-7, the residues essential for binding to CS are located on the molecular surface in the opposite side of the catalytic cleft of the protease. Therefore, it is assumed that the active site of MMP-7 bound to cell surface is directed outside. We speculate that the direction of the cell-bound MMP-7 makes it feasible for the protease to cleave its substrates on cell surface.
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Binding of Active Matrilysin to Cell Surface Cholesterol Sulfate Is Essential for Its Membrane-associated Proteolytic Action and Induction of Homotypic Cell Adhesion *
The Journal of biological chemistry, 2006Co-Authors: Kazuhiro Yamamoto, Shouichi Higashi, Mitomu Kioi, Jun Tsunezumi, Koichi Honke, Kaoru MiyazakiAbstract:Abstract Regulation of cell surface molecules by matrix metalloproteinases (MMPs), as well as MMPs-catalyzed degradation of extracellular matrix, is important for tumor invasion and metastasis. Our previous study (Kioi, M., Yamamoto, K., Higashi, S., Koshikawa, N., Fujita, K., and Miyazaki, K. (2003) Oncogene 22, 8662–8670) demonstrated that active matrilysin specifically binds to the surface of colon cancer cells and induces notable cell aggregation due to processing of the cell membrane protein(s). Furthermore, these aggregated cells showed a dramatically enhanced metastatic potential. To elucidate the mechanism of matrilysin-induced cell aggregation, we attempted to identify the matrilysin-binding substance on the cell surface. Here, we demonstrate that Cholesterol Sulfate on the cell surface is a major matrilysin-binding substance. We found that active matrilysin bound to the cell membrane and Cholesterol Sulfate incorporated into liposomes with similar affinities. Treatment of colon cancer cells with β-cyclodextrin significantly reduced not only matrilysin binding to the cell surface but also matrilysin-dependent proteolysis and cell aggregation. Interestingly, replenishment of Cholesterol Sulfate, but not Cholesterol, neutralized the effects of β-cyclodextrin. Taken together, it is likely that binding of matrilysin to Cholesterol Sulfate facilitates the matrilysin-catalyzed modulation of cell surface proteins, thus inducing the cancer cell aggregation.
Shouichi Higashi - One of the best experts on this subject based on the ideXlab platform.
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pericellular proteolysis by matrix metalloproteinase 7 is differentially modulated by Cholesterol Sulfate sulfatide and cardiolipin
FEBS Journal, 2014Co-Authors: Kazuhiro Yamamoto, Kaoru Miyazaki, Shouichi HigashiAbstract:Matrix metalloproteinase (MMP)-7 binds to cell surface Cholesterol Sulfate (CS) and acts as a membrane-associated protease. We have previously found that CS modulates the substrate preference of MMP-7, thereby regulating its pericellular proteolytic action. MMP-7 potentially associates with the cell surface via sulfatide (SM4) and cardiolipin (CL) when they are overexpressed on the cell surface. Here, we investigated the molecular interaction between these acidic lipids and MMP-7 or its substrates, and their effects on the activity of MMP-7. Studies using MMP-7 variants with low CS-binding ability suggested that these lipids interact with a similar site on MMP-7. The hydroxamate-based MMP inhibitor TAPI-1 markedly reduced the affinity of MMP-7 for CS and CL, whereas that for SM4 was not affected by TAPI-1. These three acidic lipids also had different effects on the hydrolytic activity of MMP-7 towards a small peptide substrate: SM4, CL and CS reduced the activity to 80%, 92%, and 20%, respectively. Nevertheless, SM4 and CS similarly accelerated the MMP-7-catalyzed degradation of fibronectin and laminin-332, whereas CL did not. The increased proteolysis of substrate was observed only when both substrate and enzyme had affinity for the lipid, suggesting that the lipids probably bring the reactants into closer proximity. Furthermore, MMP-7 bound to cell surface SM4 or CS cleaved specific cell surface proteins and released similar fragments, whereas the cleavage was not stimulated by cell surface CL-bound MMP-7. This study provides a novel mechanism by which acidic lipids differentially regulate pericellular proteolysis by MMP-7 through allosteric alteration of the substrate-binding site and their inherent affinities for MMP-7 substrates.
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Pericellular proteolysis by matrix metalloproteinase‐7 is differentially modulated by Cholesterol Sulfate, sulfatide, and cardiolipin
The FEBS journal, 2014Co-Authors: Kazuhiro Yamamoto, Kaoru Miyazaki, Shouichi HigashiAbstract:Matrix metalloproteinase (MMP)-7 binds to cell surface Cholesterol Sulfate (CS) and acts as a membrane-associated protease. We have previously found that CS modulates the substrate preference of MMP-7, thereby regulating its pericellular proteolytic action. MMP-7 potentially associates with the cell surface via sulfatide (SM4) and cardiolipin (CL) when they are overexpressed on the cell surface. Here, we investigated the molecular interaction between these acidic lipids and MMP-7 or its substrates, and their effects on the activity of MMP-7. Studies using MMP-7 variants with low CS-binding ability suggested that these lipids interact with a similar site on MMP-7. The hydroxamate-based MMP inhibitor TAPI-1 markedly reduced the affinity of MMP-7 for CS and CL, whereas that for SM4 was not affected by TAPI-1. These three acidic lipids also had different effects on the hydrolytic activity of MMP-7 towards a small peptide substrate: SM4, CL and CS reduced the activity to 80%, 92%, and 20%, respectively. Nevertheless, SM4 and CS similarly accelerated the MMP-7-catalyzed degradation of fibronectin and laminin-332, whereas CL did not. The increased proteolysis of substrate was observed only when both substrate and enzyme had affinity for the lipid, suggesting that the lipids probably bring the reactants into closer proximity. Furthermore, MMP-7 bound to cell surface SM4 or CS cleaved specific cell surface proteins and released similar fragments, whereas the cleavage was not stimulated by cell surface CL-bound MMP-7. This study provides a novel mechanism by which acidic lipids differentially regulate pericellular proteolysis by MMP-7 through allosteric alteration of the substrate-binding site and their inherent affinities for MMP-7 substrates.
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Identification of Amino Acid Residues of Matrix Metalloproteinase-7 Essential for Binding to Cholesterol Sulfate
The Journal of biological chemistry, 2008Co-Authors: Shouichi Higashi, Kazuhiro Yamamoto, Miwa Oeda, Kaoru MiyazakiAbstract:Matrix metalloproteinase-7 (MMP-7; matrilysin) induces homotypic adhesion of colon cancer cells by cleaving cell surface protein(s) and enhances their metastatic potential. Our previous study (Yamamoto, K., Higashi, S., Kioi, M., Tsunezumi, J., Honke, K., and Miyazaki, K. (2006) J. Biol. Chem. 281, 9170-9180) demonstrated that binding of MMP-7 to cell surface Cholesterol Sulfate (CS) is essential for the cell membrane-associated proteolytic action of the protease. To determine the region of MMP-7 essential for binding to CS, we constructed chimeric proteases consisting of various parts of MMP-7 and those of the catalytic domain of MMP-2; the latter protease does not have an affinity for CS. Studies of these chimeric proteases and other mutants of MMP-7 revealed that Ile29, Arg33, Arg51, and Trp55, in the internal sequence, and the C-terminal three residues corresponding to residues 171-173 of MMP-7 are essential for binding to CS. An MMP-7 mutant, which had the internal 4 residues at positions 29, 33, 51, and 55 of MMP-7 replaced with the corresponding residues of MMP-2 and the C-terminal 3 residues deleted, had essentially no affinity for CS. This mutant and wild-type MMP-7 showed similar proteolytic activity toward fibronectin, whereas the mutant lacked the ability to induce the colon cancer cell aggregation. In the three-dimensional structure of MMP-7, the residues essential for binding to CS are located on the molecular surface in the opposite side of the catalytic cleft of the protease. Therefore, it is assumed that the active site of MMP-7 bound to cell surface is directed outside. We speculate that the direction of the cell-bound MMP-7 makes it feasible for the protease to cleave its substrates on cell surface.
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Binding of Active Matrilysin to Cell Surface Cholesterol Sulfate Is Essential for Its Membrane-associated Proteolytic Action and Induction of Homotypic Cell Adhesion *
The Journal of biological chemistry, 2006Co-Authors: Kazuhiro Yamamoto, Shouichi Higashi, Mitomu Kioi, Jun Tsunezumi, Koichi Honke, Kaoru MiyazakiAbstract:Abstract Regulation of cell surface molecules by matrix metalloproteinases (MMPs), as well as MMPs-catalyzed degradation of extracellular matrix, is important for tumor invasion and metastasis. Our previous study (Kioi, M., Yamamoto, K., Higashi, S., Koshikawa, N., Fujita, K., and Miyazaki, K. (2003) Oncogene 22, 8662–8670) demonstrated that active matrilysin specifically binds to the surface of colon cancer cells and induces notable cell aggregation due to processing of the cell membrane protein(s). Furthermore, these aggregated cells showed a dramatically enhanced metastatic potential. To elucidate the mechanism of matrilysin-induced cell aggregation, we attempted to identify the matrilysin-binding substance on the cell surface. Here, we demonstrate that Cholesterol Sulfate on the cell surface is a major matrilysin-binding substance. We found that active matrilysin bound to the cell membrane and Cholesterol Sulfate incorporated into liposomes with similar affinities. Treatment of colon cancer cells with β-cyclodextrin significantly reduced not only matrilysin binding to the cell surface but also matrilysin-dependent proteolysis and cell aggregation. Interestingly, replenishment of Cholesterol Sulfate, but not Cholesterol, neutralized the effects of β-cyclodextrin. Taken together, it is likely that binding of matrilysin to Cholesterol Sulfate facilitates the matrilysin-catalyzed modulation of cell surface proteins, thus inducing the cancer cell aggregation.
Masao Iwamori - One of the best experts on this subject based on the ideXlab platform.
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Inhibition of cell invasion and protease activity by Cholesterol Sulfate
Fertility and sterility, 2010Co-Authors: Hanako Nakae, Hisahiko Hiroi, Mikio Momoeda, Minako Koizumi, Masao Iwamori, Yuji TaketaniAbstract:We demonstrated that Cholesterol Sulfate (CS) inhibits invasion of a trophoblast cell line and plasmin enzyme activity in a noncompetitive manner by binding to the enzyme itself, suggesting that CS can repress cell invasion by inhibiting proteinases such as those involved in the plasminogen activator/plasmin system. Considering these results, it is possible that CS may act as a signaling molecule between the trophoblast and endometrium, and may regulate the process of implantation.
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Shedding of Sulfated lipids into gastric fluid and inhibition of pancreatic DNase I by Cholesterol Sulfate in concert with bile acids.
Biochimica et Biophysica Acta, 2000Co-Authors: Masao Iwamori, Hiroko Suzuki, Takayoshi Kimura, Yuriko IwamoriAbstract:Abstract Cholesterol Sulfate (CS) and sulfatides in the epithelium of the digestive tract were found in the 1000×g supernatants of digestive fluid, particularly in gastric juices containing the duodenal contents and bile acids, there being 14–131 μg of CS and 3–54 μg of sulfatides per mg of protein in the fluid, respectively. CS and sulfatides dissolved in detergents including bile acids inactivated pancreatic trypsin to the same level as by DMSO-solubilized Sulfated lipids at 37°C. Similarly, pancreatic DNase I was inhibited by CS solubilized with DMSO or bile acids, but not by sulfatides or other membrane lipids at 37°C. Both the Sulfate group and the hydrophobic side chain of CS were indispensable structures for the inhibition of DNase I. Also, the optimum molar ratio of bile acids to CS was important for expression of the inhibitory activity of CS toward DNase I, it being 0.18 of the optimum ratio for sodium taurocholate, and the molar ratio of CS to DNase I for complete inhibition was 342:1. Thus, CS was shown to play a role as an epithelial inhibitor of DNase I in concert with bile acids.
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Alterations in Cholesterol Sulfate and its Biosynthetic Enzyme During Multistage Carcinogenesis in Mouse Skin
The Journal of investigative dermatology, 1998Co-Authors: Kaoru Kiguchi, Masao Iwamori, Ryuzaburo Higo, Miwako Kagehara, John DigiovanniAbstract:Recent evidence suggests that Cholesterol Sulfate may be an important second messenger involved in signaling epidermal differentiation in skin. The activity of Cholesterol sulfotransferase (Ch-ST) is increased during squamous differentiation of keratinocytes and is believed to be a marker enzyme for terminal differentiation. The primary objective of this study was to examine changes in levels of Cholesterol Sulfate (CS) and activity of its biosynthetic enzyme, Ch-ST, during multistage carcinogenesis in mouse skin. Using SENCAR mice, we determined the activity of Ch-ST in normal epidermis, in tumor promoter-treated epidermis, in epidermis during wound healing, and in mouse skin tumors generated by initiation-promotion regimens. A single topical application of tumor promoters led to significantly elevated levels of Ch-ST activity and of CS. Epidermal Ch-ST activity was also elevated during wound healing. Dramatic increases in CS levels and in the activity of Ch-ST were found in nearly all of the papillomas and squamous cell carcinomas examined. The increased levels of CS and activity of Ch-ST in tumor promoter-treated epidermis were accompanied by increased transglutaminase-I activity. In contrast, transglutaminase I activity was not elevated in primary papillomas or squamous cell carcinomas. Finally, Ch-ST activity was significantly elevated in the epidermis of newborn HK1.ras transgenic mice, whereas transglutaminase I activity did not correlate with Ch-ST activity in these mice. These results demonstrate that diverse tumor-promoting stimuli all produce elevated CS levels and Ch-ST activity and that CS levels and Ch-ST activity were constitutively elevated in both papillomas and squamous cell carcinomas. The data also suggest a mechanism for upregulation of Ch-ST in skin tumors involving activation/upregulation of Ha-ras.
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Alteration of acidic lipids in human sera during the course of pregnancy: Characteristic increase in the concentration of Cholesterol Sulfate
Journal of Chromatography B: Biomedical Sciences and Applications, 1997Co-Authors: Bei Lin, Yongxi Cui, Kaneyuki Kubushiro, Yasuo Akiba, Katsumi Tsukazaki, Shiro Nozawa, Masao IwamoriAbstract:In this study, we determined the concentrations of acidic lipids, including Cholesterol Sulfate (CS), sulfatide and GM3 ganglioside, in human sera of non-pregnant state and during the course of pregnancy. In human sera of non-pregnant women, GM3 was present at a concentration of 8 nmol/ml and the concentrations of CS and sulfatides were less than 20% of that of GM3. The concentration of sulfatides in sera at the second trimester of gestation was decreased, but CS gradually increased from the first to the third trimester of gestation with a correlation coefficient of 0.66, and a correlation between the concentration of CS and weeks of gestation (p
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alteration of acidic lipids in human sera during the course of pregnancy characteristic increase in the concentration of Cholesterol Sulfate
Journal of Chromatography B: Biomedical Sciences and Applications, 1997Co-Authors: Bei Lin, Yongxi Cui, Kaneyuki Kubushiro, Yasuo Akiba, Katsumi Tsukazaki, Shiro Nozawa, Masao IwamoriAbstract:In this study, we determined the concentrations of acidic lipids, including Cholesterol Sulfate (CS), sulfatide and GM3 ganglioside, in human sera of non-pregnant state and during the course of pregnancy. In human sera of non-pregnant women, GM3 was present at a concentration of 8 nmol/ml and the concentrations of CS and sulfatides were less than 20% of that of GM3. The concentration of sulfatides in sera at the second trimester of gestation was decreased, but CS gradually increased from the first to the third trimester of gestation with a correlation coefficient of 0.66, and a correlation between the concentration of CS and weeks of gestation (p<0.01). CS was also contained in the placental villi, and its concentration increased from the first to the third trimester of gestation, suggesting that placental CS is one of the source of CS in the blood by shedding.
Kenneth R. Feingold - One of the best experts on this subject based on the ideXlab platform.
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role of Cholesterol Sulfate in epidermal structure and function lessons from x linked ichthyosis
Biochimica et Biophysica Acta, 2014Co-Authors: Peter M. Elias, Mary L. Williams, Eung Ho Choi, Kenneth R. FeingoldAbstract:X-linked ichthyosis is a relatively common syndromic form of ichthyosis most often due to deletions in the gene encoding the microsomal enzyme, steroid sulfatase, located on the short area of the X chromosome. Syndromic features are mild or unapparent unless contiguous genes are affected. In normal epidermis, Cholesterol Sulfate is generated by Cholesterol sulfotransferase (SULT2B1b), but deSulfated in the outer epidermis, together forming a ‘Cholesterol Sulfate cycle’ that potently regulates epidermal differentiation, barrier function and desquamation. In XLI, Cholesterol Sulfate levels my exceed 10% of total lipid mass (≈1% of total weight). Multiple cellular and biochemical processes contribute to the pathogenesis of the barrier abnormality and scaling phenotype in XLI. This article is part of a Special Issue entitled The Important Role of Lipids in the Epidermis and their Role in the Formation and Maintenance of the Cutaneous Barrier. Guest Editors: Kenneth R. Feingold and Peter Elias.
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The mechanisms by which lipids coordinately regulate the formation of the protein and lipid domains of the stratum corneum: Role of fatty acids, oxysterols, Cholesterol Sulfate and ceramides as signaling molecules.
Dermato-endocrinology, 2011Co-Authors: Kenneth R. Feingold, Yan J. JiangAbstract:The formation of a permeability barrier between the external environment and the host is essential for survival. To provide this barrier keratinocytes undergo a complex pathway of differentiation, which culminates in keratinocyte cornification and the formation of extracellular lipid enriched lamellar membranes in the stratum corneum. The mechanisms that coordinately regulate the parallel formation of the corneocytes and lamellar membranes are unknown. The extracellular lamellar membranes are derived from the exocytosis of lamellar bodies and to synthesize lamellar bodies the keratinocyte must have abundant quantities of Cholesterol, fatty acids and ceramides. These lipids could serve as signaling molecules and thereby coordinately regulate the formation of the stratum corneum. Fatty acids activate PPARs and studies have shown that PPAR activation stimulates keratinocyte differentiation. Cholesterol is converted to oxysterols that activate LXR and studies have shown that LXR activation also stimulates keratinocyte differentation. Additionally, PPAR and LXR activation also facilitates the formation of the lipid enriched lamellar membranes. Ceramides, via a number of mechanisms also stimulate keratinocyte differentiation. Recently, studies have shown that ceramides by increasing PPAR delta also increase the expression of ABCA12, which would facilitate the formation of lamellar bodies. Finally, keratinocytes accumulate a large quantity of Cholesterol Sulfate, which plays a key role in regulating desquamation. Cholesterol Sulfate has also been shown to stimulate keratinocyte differentiation. Thus, Cholesterol, Cholesterol Sulfate, fatty acids and ceramides all stimulate keratinocyte differentiation and thereby could coordinately regulate the formation of the stratum corneum.
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Cholesterol Sulfate stimulates involucrin transcription in keratinocytes by increasing Fra-1, Fra-2, and Jun D.
Journal of lipid research, 2001Co-Authors: Karen Hanley, Peter M. Elias, Mary L. Williams, Ladonna C. Wood, Peggy Lau, Arthur H. Moser, Daniel D. Bikle, Kenneth R. FeingoldAbstract:Lipids that are synthesized de novo in the epidermis, including fatty acids, oxysterols, 1,25-dihydroxyvitamin D(3), and farnesol, can regulate the differentiation of normal human keratinocytes (NHK). Cholesterol Sulfate (CS), an epidermal lipid that is produced in the upper nucleated layers of the epidermis coincident with terminal differentiation, has been shown to play a role in the regulation of the late stages of keratinocyte differentiation, including formation of the cornified envelope. In the present study, we determined i) whether CS regulates involucrin (INV), an early keratinocyte differentiation marker, and ii) the mechanism by which CS regulates differentiation. mRNA and protein levels of INV, a precursor protein of the cornified envelope, increased 2- to 3-fold in NHK incubated in the presence of CS. In contrast, Cholesterol had no effect on INV protein or mRNA levels. Transcriptional regulation was assessed in NHK transfected with INV promoter-luciferase constructs. CS increased luciferase reporter activity approximately 2- to 3-fold in NHK transfected with a 3.7-kb INV promoter construct. Deletional analysis revealed a CS-responsive region of the INV promoter located between bp --2452 and --1880. A 5-base pair (bp) mutation of the AP-1 site (bp --2117 to --2111) within this responsive region abolished CS responsiveness, suggesting a role for the AP-1 complex in the regulation of INV transcription by CS. Electrophoretic mobility shift analysis demonstrated increased binding of nuclear extracts isolated from CS-treated NHK to AP-1 DNA as compared with vehicle-treated controls. Incubation of the nuclear extract with the appropriate antibodies showed that the AP-1 DNA-binding complex contained Fra-1, Fra-2, and Jun D. Western blots demonstrated that CS treatment increased the levels of Fra-1, Fra-2, and Jun D, and Northern analyses revealed that CS increased mRNA levels for these same AP-1 factors. These data indicate that CS, an endogenous lipid synthesized by keratinocytes, regulates the early stages of keratinocyte differentiation, and may do so through its ability to modulate levels of AP-1 proteins. -- Hanley, K., L. Wood, D. C. Ng, S. S. He, P. Lau, A. Moser, P. M. Elias, D. D. Bikle, M. L. Williams, and K. R. Feingold. Cholesterol Sulfate stimulates involucrin transcription in keratinocytes by increasing Fra-1, Fra-2, and Jun D. J. Lipid Res. 2001. 42: 390--398.
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Recessive x-Linked Ichthyosis: Role of Cholesterol-Sulfate Accumulation in the Barrier Abnormality
The Journal of investigative dermatology, 1998Co-Authors: Elizabeth Zettersten, Junko Sato, Mitsuhiro Denda, Mao-qiang Man, Angela M. Farrell, Ruby Ghadially, Mary L. Williams, Kenneth R. Feingold, Peter M. EliasAbstract:Cholesterol Sulfate is a multifunctional sterol metabolite, produced in large amounts in squamous keratinizing epithelia. Because patients with recessive x-linked ichthyosis display not only a 10-fold increase in Cholesterol Sulfate, but also a 50% reduction in Cholesterol, we assessed here whether Cholesterol Sulfate accumulation and/or Cholesterol deficiency produce abnormal barrier function in recessive x-linked ichthyosis. Patients with recessive x-linked ichthyosis display both an abnormal barrier under basal conditions, and a delay in barrier recovery after acute perturbation, which correlate with minor abnormalities in membrane structure and extensive lamellar-phase separation. Moreover, both the functional and the structural abnormalities were corrected by topical Cholesterol. Yet, topical Cholesterol Sulfate produced both a barrier abnormality in intact skin and extracellular abnormalities in isolated stratum corneum, effects largely reversed by coapplications of Cholesterol. Together, these results suggest that Cholesterol Sulfate accumulation rather than Cholesterol deficiency is responsible for the barrier abnormality. Despite the apparent importance of Cholesterol Sulfate-to-Cholesterol processing for normal barrier homeostasis, neither steroid sulfatase activity nor mRNA levels are upregulated following acute perturbations. These results demonstrate both a potential role for Cholesterol Sulfate-to-Cholesterol processing in normal permeability barrier homeostasis, and that basal levels of steroid sulfatase are sufficient to accommodate acute insults to the permeability barrier.
Toshio Kuroki - One of the best experts on this subject based on the ideXlab platform.
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Cholesterol Sulfate activates transcription of transglutaminase 1 gene in normal human keratinocytes.
The Journal of investigative dermatology, 1998Co-Authors: Shoko Kawabe, Togo Ikuta, Motoi Ohba, Kazuhiro Chida, Eichiro Ueda, Kiyofumi Yamanishi, Toshio KurokiAbstract:Cholesterol Sulfate and transglutaminase 1 are essential for the process of keratinization. Cholesterol Sulfate is formed during keratinization and activates the η isoform of protein kinase C. Transglutaminase 1 is a key enzyme for formation of the cornified envelope in terminally differentiated keratinocytes. In this study, we demonstrated that Cholesterol Sulfate acts as a transcriptional activator of the transglutaminase 1 gene in normal human keratinocytes. Growth of normal human keratinocytes was inhibited by Cholesterol Sulfate, but not by its parental Cholesterol. Treatment of normal human keratinocytes with Cholesterol Sulfate induced activity of transglutaminase 1 in a dose- and time-dependent manner. Activation of transcription of transglutaminase 1 by Cholesterol Sulfate was demonstrated by northern blotting analysis, whereas that by Cholesterol was not. In order to identify a Cholesterol Sulfate responsive region in the transglutaminase 1 gene, plasmids were constructed containing a luciferase reporter gene ligated to deletion fragments of the 5′ upstream region of the tranglutaminase 1 gene and were transfected into normal human keratinocytes. Transfected cells were treated with Cholesterol Sulfate, the phorbol ester 12-O-tetradecanoylphorbol-13-acetate and a high concentration of Ca2+. Our results indicate that the responsive element(s) for Cholesterol Sulfate and phorbol ester is located upstream of the human transglutaminase 1 gene at a position(s) between –819 and –549, whereas the responsive element for Ca2+ is located at a position between –79 and –49.
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Cholesterol Sulfate, a second messenger for the η isoform of protein kinase C, inhibits promotional phase in mouse skin carcinogenesis
Cancer research, 1995Co-Authors: Kazuhiro Chida, Tohgo Ikuta, Akiko Murakami, Tomoko Tagawa, Toshio KurokiAbstract:Abstract Cholesterol Sulfate is a second messenger for the η isoform of protein kinase C mediating squamous differentiation. We found that Cholesterol Sulfate inhibited the promotional phase of skin carcinogenesis in female CD-1 mice, which was initiated by 100 µg 7,12-dimethylbenz[ a ]-anthracene and promoted by a single application of 10 µg 12- O -tetradecanoylphorbol-13-acetate, followed by repeated applications of 10 µg mezerein once a week for 19 weeks. Cholesterol Sulfate, when applied topically at a dose of 400 µg (820 µmol) 10 min before treatment with the promoters, markedly suppressed tumor formation, resulting in decrease of 56% in the incidence of tumor-bearing mice, 81% in the number of tumors/mouse, and 60% in the size of tumors at 20 weeks of the promotion. This inhibition was not due to elimination of the initiated cells. Treatment with the parental Cholesterol at a dose of 320 µg (820 µmol), which does not activate the η isoform, did not inhibit tumor promotion. Repeated treatment with Cholesterol Sulfate induced scaling of skin at the site of application. Cholesterol Sulfate, unlike most inhibitors of tumor promotion, did not inhibit induction of ornithine decarboxylase and hyperplasia in mouse epidermis caused by topical treatment with 12- O -tetradecanoylphorbol-13-acetate. These findings suggest that Cholesterol Sulfate inhibits tumor promotion by stimulating a differentiation pathway mediated by the η isoform of protein kinase C.
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Cholesterol Sulfate, a novel activator for the eta isoform of protein kinase C.
Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research, 1994Co-Authors: Tohgo Ikuta, Kazuhiro Chida, O Tajima, Y Matsuura, M Iwamori, Y Ueda, Keiko Mizuno, Shigeo Ohno, Toshio KurokiAbstract:Activity of protein kinase C depends on the interaction with polar head-groups of two membrane lipids, i.e., phosphatidylserine and diacylglycerol. In the present study, we demonstrated a novel activation mechanism of the eta isoform of protein kinase C (nPKC eta), which is predominantly expressed in epithelial tissues in close association with epithelial differentiation. We found that the nPKC eta was activated by Cholesterol Sulfate, a metabolite of Cholesterol formed during squamous differentiation. This activation was greater than that by phosphatidylserine plus phorbol ester; the Vmax for the activation by Cholesterol Sulfate was 3.6 times that by phosphatidylserine plus phorbol ester, while Kms were almost equal. In the presence of Cholesterol Sulfate, phorbol ester only weakly enhanced the activity of nPKC eta. Activation of nPKC eta by Cholesterol Sulfate was further demonstrated by autophosphorylation of the kinase molecule. However, the alpha and delta isoforms of protein kinase C were not activated by Cholesterol Sulfate. The present observation affords a new insight into a signal transduction pathway of squamous differentiation.