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PMID: 236354 Published · ppublish English Journal Article

Subcellular localization and heterogeneity of neutral proteases in neutrophilic polymorphonuclear leukocytes.

The Journal of experimental medicine ·Vol. 141 ·No. 4 ·1975-04-01 ·Pages 709-23

Dewald B, Rindler-Ludwig R, Bretz U, Baggiolini M

Abstract

The subcellular localization of elastase and of neutral proteases hydrolyzing histone and casein was determined in human and rabbit polymorphonuclear leukocytes using fractionation by isopycnic centrifugation. Granule-rich fractions obtained by this technique were extracted and analyzed by acrylamide gel electrophoresis, and proteolytic activity on the gels was demonstrated by staining with either N-acetyl-D,L-alanine alpha-naphthyl ester or naphthol AS-D acetate as substrate. In both species, all neutral proteases assayed were found to be localized exclusively in the azurophil granules. Specific activities were about 10-30 times higher in human than in rabbit preparations. In extracts of human azurophil granules up to 10 proteins exhibiting esterolytic activity could be demonstrated after electrophoretic separation. Three major and two or three minor components of these esterases were shown to possess elastase activity. Similar zymograms prepared with extracts from rabbit azurophil granules revealed only one major elastase band. The electrophoretic analysis further showed that the most strongly cationic proteins of both human and rabbit PMNs were also confined to the azurophil granules.

MeSH Terms
Alkaline Phosphatase/analysis Animals Caseins Catalysis Centrifugation, Isopycnic Cytoplasmic Granules/enzymology Electrophoresis, Polyacrylamide Gel Hexosaminidases/analysis Histones Humans Hydrogen-Ion Concentration Muramidase/analysis Neutrophils/enzymology,ultrastructure Pancreatic Elastase/analysis Peptide Hydrolases/analysis Peroxidases/analysis Rabbits Subcellular Fractions/enzymology Time Factors
Chemicals
Caseins Histones Peroxidases Alkaline Phosphatase Hexosaminidases Muramidase Peptide Hydrolases Pancreatic Elastase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Dewald B
Rindler-Ludwig R
Bretz U
Baggiolini M
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43 references, click to expand
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Article Info
Journal
The Journal of experimental medicine
Abbr.
J Exp Med
ISSN
0022-1007
DOI
10.1084/jem.141.4.709
Published
1975-04-01
Pages
709-23
Language
English
Region
United States
NLM ID
2985109R
PMCID
PMC2189758
Subset
IM
External Links
PubMed source DOI: 10.1084/jem.141.4.709 PMC Full Text View journal details
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