Collagen V Is a Potential Substrate for Clostridial Collagenase G in Pancreatic Islet Isolation

  • J Diabetes Res. 2016:2016:4396756. doi: 10.1155/2016/4396756.
Hiroki Shima  1 ,  Akiko Inagaki  2 ,  Takehiro Imura  3 ,  Youhei Yamagata  4 ,  Kimiko Watanabe  5 ,  Kazuhiko Igarashi  6 ,  Masafumi Goto  2 ,  Kazutaka Murayama  7
Affiliations
  • 1. Department of Biochemistry, Tohoku University Graduate School of Medicine, Sendai 980-8575, Japan; AMED-CREST, Japan Agency for Medical Research and Development, Tokyo 100-0004, Japan.
  • 2. Division of Transplantation and Regenerative Medicine, Tohoku University School of Medicine, Sendai 980-8575, Japan.
  • 3. Division of Transplantation and Regenerative Medicine, Tohoku University School of Medicine, Sendai 980-8575, Japan; New Industry Creation Hatchery Center, Tohoku University, Sendai 980-8579, Japan.
  • 4. Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, Fuchu 183-8509, Japan.
  • 5. New Industry Creation Hatchery Center, Tohoku University, Sendai 980-8579, Japan.
  • 6. Department of Biochemistry, Tohoku University Graduate School of Medicine, Sendai 980-8575, Japan; AMED-CREST, Japan Agency for Medical Research and Development, Tokyo 100-0004, Japan; Center for Regulatory Epigenome and Diseases, Tohoku University Graduate School of Medicine, Sendai 980-8575, Japan.
  • 7. Graduate School of Biomedical Engineering, Tohoku University, Sendai 980-8575, Japan.
PMID: 27195301 DOI: 10.1155/2016/4396756
Abstract

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.

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