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