A simple two-step purification of protease nexin.

David Farrell, W. E. Van Nostrand, D. D. Cunningham

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

This paper describes a simple purification procedure for protease nexin, a serine proteinase inhibitor secreted by cultured human fibroblasts that regulates proteinase activity at and near the cell surface. The first step in the procedure takes advantage of the high-affinity binding of protease nexin to dextran sulphate-Sepharose. This step eliminates the need for prior concentration of the serum-free fibroblast-conditioned medium, since protease nexin binds to the resin in the presence of physiological saline. The use of dextran sulphate also provides an affinity resin with considerably less variability than the heparin-based resins previously used. Final purification to homogeneity involves a combination of DEAE-Sepharose in-line with dextran sulphate-Sepharose to simultaneously purify and concentrate the protein. Purified protease nexin is shown by Ouchterlony analysis and peptide mapping to be immunologically and structurally distinct from antithrombin III and heparin cofactor II, two plasma proteinase inhibitors with similar properties.

Original languageEnglish (US)
Pages (from-to)907-912
Number of pages6
JournalBiochemical Journal
Volume237
Issue number3
StatePublished - Aug 1 1986
Externally publishedYes

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Protease Nexins
Dextran Sulfate
Purification
Sepharose
Resins
Fibroblasts
Peptide Hydrolases
Heparin Cofactor II
Serine Proteinase Inhibitors
Peptide Mapping
Antithrombin III
Conditioned Culture Medium
Heparin
Plasmas
Peptides
Serum
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Farrell, D., Van Nostrand, W. E., & Cunningham, D. D. (1986). A simple two-step purification of protease nexin. Biochemical Journal, 237(3), 907-912.

A simple two-step purification of protease nexin. / Farrell, David; Van Nostrand, W. E.; Cunningham, D. D.

In: Biochemical Journal, Vol. 237, No. 3, 01.08.1986, p. 907-912.

Research output: Contribution to journalArticle

Farrell, D, Van Nostrand, WE & Cunningham, DD 1986, 'A simple two-step purification of protease nexin.', Biochemical Journal, vol. 237, no. 3, pp. 907-912.
Farrell D, Van Nostrand WE, Cunningham DD. A simple two-step purification of protease nexin. Biochemical Journal. 1986 Aug 1;237(3):907-912.
Farrell, David ; Van Nostrand, W. E. ; Cunningham, D. D. / A simple two-step purification of protease nexin. In: Biochemical Journal. 1986 ; Vol. 237, No. 3. pp. 907-912.
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