Document Detail

Purification and sequence analysis of two rat tissue inhibitors of metalloproteinases.
MedLine Citation:
PMID:  1309971     Owner:  NLM     Status:  MEDLINE    
Two protein inhibitors of metalloproteinases (TIMP) were isolated from medium conditioned by the clonal rat osteosarcoma line UMR 106-01. Initial purification of both a 30-kDa inhibitor and a 20-kDa inhibitor was accomplished using heparin-Sepharose chromatography with dextran sulfate elution followed by DEAE-Sepharose and CM-Sepharose chromatography. Purification of the 20-kDa inhibitor to homogeneity was completed with reverse-phase high-performance liquid chromatography. The 20-kDa inhibitor was identified as rat TIMP-2. The 30-kDa inhibitor, although not purified to homogeneity, was identified as rat TIMP-1. Amino terminal amino acid sequence analysis of the 30-kDa inhibitor demonstrated 86% identity to human TIMP-1 for the first 22 amino acids while the sequence of the 20-kDa inhibitor was identical to that of human TIMP-2 for the first 22 residues. Treatment with peptide:N-glycosidase F indicated that the 30-kDa rat inhibitor is glycosylated while the 20-kDa inhibitor is apparently unglycosylated. Inhibition of both rat and human interstitial collagenase by rat TIMP-2 was stoichiometric, with a 1:1 molar ratio required for complete inhibition. Exposure of UMR 106-01 cells to 10(-7) M parathyroid hormone resulted in approximately a 40% increase in total inhibitor production over basal levels.
W T Roswit; D W McCourt; N C Partridge; J J Jeffrey
Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Archives of biochemistry and biophysics     Volume:  292     ISSN:  0003-9861     ISO Abbreviation:  Arch. Biochem. Biophys.     Publication Date:  1992 Feb 
Date Detail:
Created Date:  1992-02-18     Completed Date:  1992-02-18     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0372430     Medline TA:  Arch Biochem Biophys     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  402-10     Citation Subset:  IM; S    
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63130.
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MeSH Terms
Amino Acid Sequence
Cell Line
Chromatography, Affinity
Chromatography, High Pressure Liquid
Chromatography, Ion Exchange
Electrophoresis, Polyacrylamide Gel
Glycoproteins / chemistry,  isolation & purification*,  pharmacology
Microbial Collagenase / antagonists & inhibitors
Molecular Sequence Data
Molecular Weight
Parathyroid Hormone / pharmacology
Sequence Homology, Nucleic Acid
Tissue Inhibitor of Metalloproteinases
Grant Support
Reg. No./Substance:
0/Glycoproteins; 0/Parathyroid Hormone; 0/Tissue Inhibitor of Metalloproteinases; EC Collagenase
N C Partridge / St Louis Sch Med, MO

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