Glycosaminoglycans on fibroblasts accelerate thrombin inhibition by protease nexin-1.

David Farrell, D. D. Cunningham

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Protease nexin-1 (PN-1) is a proteinase inhibitor that is secreted by human fibroblasts in culture. PN-1 inhibits certain regulatory serine proteinases by forming a covalent complex with the catalytic-site serine residue; the complex then binds to the cell surface and is internalized and degraded. The fibroblast surface was recently shown to accelerate the rate of complex-formation between PN-1 and thrombin. The present paper demonstrates that the accelerative activity is primarily due to cell-surface heparan sulphate, with a much smaller contribution from chondroitin sulphate. This conclusion is supported by the effects of purified glycosaminoglycans on the second-order rate constant for the inhibition of thrombin by PN-1. Also, treatment of 35SO4(2-)-labelled cells with heparitin sulphate lyase or chondroitin sulphate ABC lyase demonstrated two discrete pools of 35S-labelled glycosaminoglycans; subsequent treatment of plasma membranes with these glycosidases showed that heparitin sulphate lyase treatment abolished about 80% of the accelerative activity and chondroitin sulphate ABC lyase removed the remaining 20%. These results show that two components are responsible for the acceleration of PN-1-thrombin complex-formation by human fibroblasts. Although dermatan sulphate is also present on fibroblasts, it did not accelerate the inhibition of thrombin by PN-1.

Original languageEnglish (US)
Pages (from-to)543-550
Number of pages8
JournalBiochemical Journal
Volume245
Issue number2
StatePublished - Jul 15 1987
Externally publishedYes

Fingerprint

Protease Nexins
Fibroblasts
Glycosaminoglycans
Thrombin
Lyases
Heparitin Sulfate
Chondroitin Sulfates
Chondroitin Lyases
Chondroitin ABC Lyase
Dermatan Sulfate
Glycoside Hydrolases
Serine Proteases
Cell membranes
Cell culture
Serine
Rate constants
Catalytic Domain
Peptide Hydrolases
Cell Membrane

ASJC Scopus subject areas

  • Biochemistry

Cite this

Glycosaminoglycans on fibroblasts accelerate thrombin inhibition by protease nexin-1. / Farrell, David; Cunningham, D. D.

In: Biochemical Journal, Vol. 245, No. 2, 15.07.1987, p. 543-550.

Research output: Contribution to journalArticle

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