A salt linkage between phe-88 and asp-432 is responsible for the ca++ -independent activity of gelatinase

B. Chun, Hui Bu, Tayebeh Pourmotabbed

Research output: Contribution to journalArticle

Abstract

Progelatinase B can be activated in vitro by organomercurial compounds and by proteolytic enzymes such as trypsin, chymotrypsin and stromelysin. Activation of the proenzyme by either 4-aminophenylmercuric acetate (APMA) or chymotrypsin yielded proteins that absolutely required Ca++ for activity regardless of pH of the reaction mixture. The trypsin- and stromelysin-activated gelatinases, on the other hand, did not require Ca++ for activity at pH 7.5, but the activity of the trypsin-activated enzyme became Ca++ -dependent as pH increased. The pH study revealed that an amino acid residue with an apparent pÄ' of 8.8 was involved in this process. The amino-terminal analyses showed that trypsin- and stromelysin-activated enzymes had the same amino termini (Phe-88), but APMA- and chymotrypsin-activated enzymes had Met-75 and Gin-89 or Glu-92 as the amino-terminal amino acid, respectively. Replacing Asp-432 in progelatinase with either Glu, Asn, Gly, or Lys resulted in the proteins that, upon activation by trypsin required Ca++ for activity. These data, in conjunction with the X-ray crystal structure of collagenase, suggest that a salt linkage between Phe-88 and Asp-432 is able to mimic the role of Ca++ in catalysis of gelatinase B, and is responsible for the Ca++-independent activity of the trypsin- and stromelysin-activated enzyme.

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number6
StatePublished - Dec 1 1996

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Gelatinases
Matrix Metalloproteinase 3
Trypsin
Salts
Chymotrypsin
Enzymes
Chemical activation
Amino Acids
Enzyme Precursors
Matrix Metalloproteinase 9
Collagenases
Catalysis
Proteins
Peptide Hydrolases
Crystal structure
X-Rays
X rays

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

A salt linkage between phe-88 and asp-432 is responsible for the ca++ -independent activity of gelatinase. / Chun, B.; Bu, Hui; Pourmotabbed, Tayebeh.

In: FASEB Journal, Vol. 10, No. 6, 01.12.1996.

Research output: Contribution to journalArticle

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AU - Pourmotabbed, Tayebeh

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