Collagen V Is a Potential Substrate for Clostridial Collagenase G in Pancreatic Islet Isolation
The clostridial collagenases, H and G, play key roles in pancreatic islet isolation. Collagenases digest the peptide bond between Yaa and the subsequent Gly in Gly-Xaa-Yaa repeats. To fully understand the pancreatic islet isolation process, identification of the collagenase substrates in the tissue...
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| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2016-01-01
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| Series: | Journal of Diabetes Research |
| Online Access: | http://dx.doi.org/10.1155/2016/4396756 |
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| _version_ | 1832563599383461888 |
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| author | Hiroki Shima Akiko Inagaki Takehiro Imura Youhei Yamagata Kimiko Watanabe Kazuhiko Igarashi Masafumi Goto Kazutaka Murayama |
| author_facet | Hiroki Shima Akiko Inagaki Takehiro Imura Youhei Yamagata Kimiko Watanabe Kazuhiko Igarashi Masafumi Goto Kazutaka Murayama |
| author_sort | Hiroki Shima |
| collection | DOAJ |
| description | The clostridial collagenases, H and G, play key roles in pancreatic islet isolation. Collagenases digest the peptide bond between Yaa and the subsequent Gly in Gly-Xaa-Yaa repeats. To fully understand the pancreatic islet isolation process, identification of the collagenase substrates in the tissue is very important. Although collagen types I and III were reported as possible substrates for collagenase H, the substrate for collagenase G remains unknown. In this study, collagen type V was focused upon as the target for collagenases. In vitro digestion experiments for collagen type V were performed and analyzed by SDS-PAGE and mass spectrometry. Porcine pancreatic tissues were digested in vitro under three conditions and observed during digestion. The results revealed that collagen type V was only digested by collagenase G and that the digestion was initiated from the N-terminal part. Tissue degradation during porcine islet isolation was only observed in the presence of both collagenases H and G. These findings suggest that collagen type V is one of the substrates for collagenase G. The enzymatic activity of collagenase G appears to be more important for pancreatic islet isolation in large mammals such as pigs and humans. |
| format | Article |
| id | doaj-art-e1da7bd797bb4ab0b48d0926cc76f72c |
| institution | Kabale University |
| issn | 2314-6745 2314-6753 |
| language | English |
| publishDate | 2016-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Diabetes Research |
| spelling | doaj-art-e1da7bd797bb4ab0b48d0926cc76f72c2025-02-03T01:13:14ZengWileyJournal of Diabetes Research2314-67452314-67532016-01-01201610.1155/2016/43967564396756Collagen V Is a Potential Substrate for Clostridial Collagenase G in Pancreatic Islet IsolationHiroki Shima0Akiko Inagaki1Takehiro Imura2Youhei Yamagata3Kimiko Watanabe4Kazuhiko Igarashi5Masafumi Goto6Kazutaka Murayama7Department of Biochemistry, Tohoku University Graduate School of Medicine, Sendai 980-8575, JapanDivision of Transplantation and Regenerative Medicine, Tohoku University School of Medicine, Sendai 980-8575, JapanDivision of Transplantation and Regenerative Medicine, Tohoku University School of Medicine, Sendai 980-8575, JapanGraduate School of Agricultural Science, Tokyo University of Agriculture and Technology, Fuchu 183-8509, JapanNew Industry Creation Hatchery Center, Tohoku University, Sendai 980-8579, JapanDepartment of Biochemistry, Tohoku University Graduate School of Medicine, Sendai 980-8575, JapanDivision of Transplantation and Regenerative Medicine, Tohoku University School of Medicine, Sendai 980-8575, JapanGraduate School of Biomedical Engineering, Tohoku University, Sendai 980-8575, JapanThe clostridial collagenases, H and G, play key roles in pancreatic islet isolation. Collagenases digest the peptide bond between Yaa and the subsequent Gly in Gly-Xaa-Yaa repeats. To fully understand the pancreatic islet isolation process, identification of the collagenase substrates in the tissue is very important. Although collagen types I and III were reported as possible substrates for collagenase H, the substrate for collagenase G remains unknown. In this study, collagen type V was focused upon as the target for collagenases. In vitro digestion experiments for collagen type V were performed and analyzed by SDS-PAGE and mass spectrometry. Porcine pancreatic tissues were digested in vitro under three conditions and observed during digestion. The results revealed that collagen type V was only digested by collagenase G and that the digestion was initiated from the N-terminal part. Tissue degradation during porcine islet isolation was only observed in the presence of both collagenases H and G. These findings suggest that collagen type V is one of the substrates for collagenase G. The enzymatic activity of collagenase G appears to be more important for pancreatic islet isolation in large mammals such as pigs and humans.http://dx.doi.org/10.1155/2016/4396756 |
| spellingShingle | Hiroki Shima Akiko Inagaki Takehiro Imura Youhei Yamagata Kimiko Watanabe Kazuhiko Igarashi Masafumi Goto Kazutaka Murayama Collagen V Is a Potential Substrate for Clostridial Collagenase G in Pancreatic Islet Isolation Journal of Diabetes Research |
| title | Collagen V Is a Potential Substrate for Clostridial Collagenase G in Pancreatic Islet Isolation |
| title_full | Collagen V Is a Potential Substrate for Clostridial Collagenase G in Pancreatic Islet Isolation |
| title_fullStr | Collagen V Is a Potential Substrate for Clostridial Collagenase G in Pancreatic Islet Isolation |
| title_full_unstemmed | Collagen V Is a Potential Substrate for Clostridial Collagenase G in Pancreatic Islet Isolation |
| title_short | Collagen V Is a Potential Substrate for Clostridial Collagenase G in Pancreatic Islet Isolation |
| title_sort | collagen v is a potential substrate for clostridial collagenase g in pancreatic islet isolation |
| url | http://dx.doi.org/10.1155/2016/4396756 |
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