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Methods for Assessments of Collagenolytic Activity of the Vibrio cholerae Extracellular Proteases, Purification of Secreted Collagenase VchC, and Extraction of Type I Collagen from Fish Skin

  • Bo R. Weber
  • Aleksandra E. SikoraEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1839)

Abstract

Secreted proteases have been linked to facilitating the survival of Vibrio cholerae in different environmental niches. Examination of protease activity using various measures is critical to determine the substrate specificity of a given proteolytic enzyme and its biological function. Here, we describe methods to assess collagenolytic activity of the V. cholerae extracellular proteases using synthetic and natural substrates in quantitative and qualitative assays. Purification of the secreted V. cholerae collagenase VchC and extraction of type I collagen from fish skin are also described.

Key words

Metalloprotease Collagenolytic proteases Secretion Collagenase Type I collagen SDS-PAGE FALGPA Zymography Affinity chromatography DQ-gelatin 

Notes

Acknowledgments

This work was supported by the start-up funds from Oregon State University (OSU) as well as Faculty Development Funding to AES.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Pharmaceutical Sciences, College of PharmacyOregon State UniversityCorvallisUSA

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