Purification and characterization of a human neutrophil neutral protease. The neutral peptide-generating protease

The Journal of Clinical Investigation
J S CoblynB U Wintroub

Abstract

A human neutrophil neutral protease which generates a low molecular weight peptide from a plasma protein substrate and cleaves the basic amino acid ester substrates alpha-N-p-tosyl-l-arginine methyl ester HCl, alpha-N-benzoyl-l-arginine-methyl ester HCl, and alpha-N-carbobenzoxy-l-lysine-p-nitrophenyl ester has been purified to homogeneity and distinguished from the known lysosomal neutrophil proteases. The starting activity was obtained from purified human neutrophils by homogenization, sedimentation by low-speed centrifugation, and high salt elution of the insoluble material. Purification was achieved by aprotinin-affinity chromatography, precipitation at low ionic strength, and gel filtration. The overall recovery, relative to the activity in the starting eluate of the neutrophil fraction, was congruent with50% with a 200- to 400-fold increase in specific activity. After treatment with diisopropylfluorophosphate to eliminate autodegradation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of reduced and unreduced protein gave a single protein band of 29,000-30,000 mol wt. The isoelectric point determined in sucrose gradients ranged from pH 7.8 to 8.3 with a peak at pH 8.0. This neutrophil protease, like cathepsin G...Continue Reading

References

Sep 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·B U WintroubK F Austen
Feb 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·B U WintroubK F Austen
Mar 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·S PontremoliB L Horecker
Jan 1, 1982·The Journal of Clinical Investigation·M G TonnesenB U Wintroub

Citations

Feb 27, 1975·Biochimica Et Biophysica Acta·R Rindler-Ludwig, H Braunsteiner
Jul 1, 1975·The Journal of Clinical Investigation·E F Plow, T S Edgington
Jan 11, 1977·Biochimica Et Biophysica Acta·M Zimmerman, B M Ashe
Jun 2, 1971·Nature: New Biology·G WeissmannR B Zurier
Apr 7, 1972·Biochimica Et Biophysica Acta·L Visser, E R Blout
Nov 1, 1955·Proceedings of the Society for Experimental Biology and Medicine·L A SACHARS FRANKEL
Dec 1, 1959·Canadian Journal of Biochemistry and Physiology·B C HUMMEL

Related Concepts

Neutrophil Band Cells
Elastases
Cathepsin G
Neutral peptide-generating protease
Isoflurophate
SDS-PAGE
Peptide Hydrolases
Neutrophils as Percentage of Blood Leukocytes (Lab Test)
Serum Proteins
Plasma Protein Assay

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