PMID: 2121162Jul 1, 1990Paper

Monoclonal antibodies specific for neutrophil proteinase 4. Production and use for isolation of the enzyme

Biological Chemistry Hoppe-Seyler
K OhlssonM Rosengren

Abstract

Four stable hybridoma cell lines producing monoclonal antibodies specific for neutrophil proteinase 4 (NP4) were established and one monoclonal antibody was chosen to produce an immunoaffinity-resin for the purification of NP4. In a precipitation assay system these antibodies bound NP4 in a dose-dependent manner, but did so neither with neutrophil elastase nor with cathepsin G. NP4 was purified and electrophoresis of the affinity-purified enzyme in sodium dodecyl sulfate polyacrylamide gels resulted in a single Mr = 30,000 polypeptide. The purified enzyme digested fibrin but not elastin and it cleaved Boc-Ala-ONp readily (Km = 0.47mM) at neutral pH, but had no effect on Suc-[Ala]3 Nan and N-Suc-[Ala]2-Pro-Phe-pNA. The proteolytic activity was inhibited by DFP, alpha 1 PI and alpha 2 M with a Ki of 10(-9)M for the NP4-alpha 1 PI complex. The NH2-terminal sequence and the amino-acid composition of NP4 were distinct from those of elastase and cathepsin G. Neutrophils contain large amounts of NP4 as judged by the comparable amounts of elastase- and NP4-alpha 1 PI complexes present in inflammatory exudates.

References

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Citations

Apr 27, 2013·Neurological Sciences : Official Journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology·Xiaoye MoZhiping Hu
Sep 1, 1995·General Pharmacology·P Renesto, M Chignard
May 1, 1997·Atherosclerosis·A M ElneihoumK Ohlsson
Jun 1, 1993·American Journal of Respiratory Cell and Molecular Biology·N V RaoJ R Hoidal
Apr 1, 1994·Scandinavian Journal of Gastroenterology·M BergenfeldtK Ohlsson
Nov 1, 1995·Scandinavian Journal of Clinical and Laboratory Investigation·L AxelssonK Ohlsson
Dec 1, 1992·Scandinavian Journal of Clinical and Laboratory Investigation·M BergenfeldtK Ohlsson
Dec 31, 1992·Annals of the New York Academy of Sciences·A J Barrett
Jan 1, 1991·Annals of the New York Academy of Sciences·P DaviesR A Mumford
Oct 1, 1993·Gastroenterologia Japonica·M Bergenfeldt, K Ohlsson
May 1, 1992·Biological Chemistry Hoppe-Seyler·M BergenfeldtK Ohlsson

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