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PMID: 2039471 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Purification of recombinant human prostromelysin. Studies on heat activation to give high-Mr and low-Mr active forms, and a comparison of recombinant with natural stromelysin activities.

The Biochemical journal ·Vol. 276 ( Pt 1) ·1991-05-15 ·Pages 217-21

Koklitis PA, Murphy G, Sutton C, Angal S

Abstract

Recombinant human prostromelysin was purified in a single step using Procion Red-Sepharose chromatography. The purified prostromelysin was self-activated to high-Mr (45,000) and low-Mr (28,000) forms by incubation at 55 degrees C without the addition of extraneous activators. The two forms of stromelysin were subsequently separated, again using Procion Red-Sepharose. Both of the heat-activated recombinant forms demonstrated similar specific activities (for the macromolecular substrates casein, gelatin, elastin, proteoglycan and type IV collagen) when compared with either heat- or trypsin-activated natural stromelysin. The heat-activated recombinant stromelysins both showed similar abilities to potentiate activation of human procollagenase when compared with trypsin-activated natural stromelysin.

MeSH Terms

Amino Acid Sequence Cells, Cultured Chromatography, Affinity/methods Electrophoresis, Polyacrylamide Gel Enzyme Activation Enzyme Precursors/genetics,isolation & purification,metabolism Fibroblasts/enzymology Gingiva/enzymology Glycoproteins/isolation & purification,pharmacology Hot Temperature Humans Kinetics Matrix Metalloproteinase 3 Metalloendopeptidases/genetics,isolation & purification,metabolism Molecular Sequence Data Molecular Weight Peptide Fragments/isolation & purification Recombinant Proteins/isolation & purification,metabolism,pharmacology Tissue Inhibitor of Metalloproteinases

Chemicals

Enzyme Precursors Glycoproteins Peptide Fragments Recombinant Proteins Tissue Inhibitor of Metalloproteinases Metalloendopeptidases prostromelysin Matrix Metalloproteinase 3

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Koklitis P A

Celltech Limited, Slough, U.K.

Murphy G

Sutton C

Angal S

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Article Info

Journal

The Biochemical journal

Abbr.

Biochem J

ISSN

0264-6021

DOI

10.1042/bj2760217

Published

1991-05-15

Pages

217-21

Language

English

Region

England

NLM ID

2984726R

PMCID

PMC1151167

Subset

External Links

PubMed source DOI: 10.1042/bj2760217 PMC Full Text View journal details

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Purification of recombinant human prostromelysin. Studies on heat activation to give high-Mr and low-Mr active forms, and a comparison of recombinant with natural stromelysin activities.

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