Document Detail


Different roles of class I and class II Clostridium histolyticum collagenase in rat pancreatic islet isolation.
MedLine Citation:
PMID:  7859945     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Crude Clostridium histolyticum collagenase was purified by gel filtration and fractionated by anion exchange chromatography into class I with high collagen digestion activity (CDA) and low FALGPA (2-furanacryloyl-L-leucylglycyl-L-prolyl-L-alanine) hydrolysis activity (FHA), class II with low CDA and high FHA, and a fraction called class I/II with intermediate activities. The roles of these collagenase classes in rat pancreatic islet isolation were investigated. Dissociations were carried out with 360 mg of pancreatic tissue in 10 ml of buffer containing 10% (wt/vol) albumin to suppress endogenous proteolytic activity, 100 U of C. histolyticum neutral protease, and one or two purified collagenase(s). For purified nonfractionated (PNF) collagenase, 2.6 mg of enzyme containing 2.4 U CDA and 38.0 U FHA was used, and for the separate classes, comparable amounts of activity were added. PNF collagenase dissociated the tissue completely in 32 min and yielded 5.0 +/- 0.4 microliters islet tissue/g pancreas. Class I collagenase alone dissociated pancreatic tissue extremely slowly and incompletely; only a few islets were released (0.7 +/- 0.2 microliters/g pancreas). Class II collagenase alone dissociated the tissue adequately in 50 min, and a high islet yield of 5.7 +/- 0.6 microliters/g was obtained. With class I/II, a similar dissociation time (47 min) and islet yield (5.5 +/- 0.3 microliters/g) were obtained. Combining class I and class II collagenase resulted in a more rapid dissociation (32 min) and a higher islet yield (7.1 +/- 0.8 microliters/g) than that obtained with PNF collagenase (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)
Authors:
G H Wolters; G H Vos-Scheperkeuter; H C Lin; R van Schilfgaarde
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Diabetes     Volume:  44     ISSN:  0012-1797     ISO Abbreviation:  Diabetes     Publication Date:  1995 Feb 
Date Detail:
Created Date:  1995-03-21     Completed Date:  1995-03-21     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0372763     Medline TA:  Diabetes     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  227-33     Citation Subset:  AIM; IM    
Affiliation:
Department of Surgery, University of Groningen, The Netherlands.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Cell Separation / methods*
Chromatography, Gel
Chromatography, Ion Exchange
Islets of Langerhans / cytology*,  physiology
Male
Microbial Collagenase / isolation & purification,  metabolism*
Molecular Sequence Data
Rats
Rats, Wistar
Chemical
Reg. No./Substance:
EC 3.4.24.3/Microbial Collagenase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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