Degradation of human anaphylatoxin C3a by rat peritoneal mast cells: a role for the secretory granule enzyme chymase and heparin proteoglycan
Purified human C3a was iodinated (/sup 125/I-C3a) and used to study the interaction of labeled peptide with rat peritoneal mast cells (RMC). Cellular binding of /sup 125/I-C3a occurred within 30 sec, followed by a rapid dissociation from the cell. Once /sup 125/I-C3a was exposed to RMC, it lost the ability to rebind to a second batch of RMC. Analysis of the supernatants by trichloroacetic acid (TCA) precipitation and electrophoresis in sodium dodecyl sulfate polyacrylamide gels (SDS PAGE) revealed a decrease in the fraction of /sup 125/I precipitable by TCA and the appearance of /sup 125/-C3a cleavage fragments. Pretreatment of RMC with enzyme inhibitors specific for chymotrypsin, but not trypsin, abrogated the degradation of /sup 125/I-C3a. Treatment of RMC bearing /sup 125/I-C3a with bis (sulfosuccinimidyl) suberate (BS/sup 3/) covalently cross-linked the /sup 125/I-C3a to chymase, the predominant enzyme found in the secretory granules. Indirect immunofluorescence of RMC by using the IgG fraction of goat anti-rat chymase showed that chymase is present on the surface of unstimulated cells. Neither purified chymase nor heparin proteoglycan alone had any appreciable effect on /sup 125/I-C3a, but together they resulted in prompt degradation of the /sup 125/I-C3a.
- Research Organization:
- Medical College of Virginia, Richmond
- OSTI ID:
- 7092132
- Journal Information:
- J. Immunol.; (United States), Vol. 136:1
- Country of Publication:
- United States
- Language:
- English
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SERINE PROTEINASES
BIOCHEMICAL REACTION KINETICS
ANAPHYLAXIS
ELECTROPHORESIS
ENZYMATIC HYDROLYSIS
GLUCOPROTEINS
HEPARIN
IODINE 125
LABELLED COMPOUNDS
MAST CELLS
RADIORECEPTOR ASSAY
RATS
RECEPTORS
AMINES
ANIMAL CELLS
ANIMALS
ANTICOAGULANTS
BETA DECAY RADIOISOTOPES
CARBOHYDRATES
CHEMICAL REACTIONS
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DECOMPOSITION
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
ENZYMES
HEMATOLOGIC AGENTS
HYDROLASES
HYDROLYSIS
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LYSIS
MAMMALS
MEMBRANE PROTEINS
MUCOPOLYSACCHARIDES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PEPTIDE HYDROLASES
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RADIOISOTOPES
REACTION KINETICS
RODENTS
SACCHARIDES
SOLVOLYSIS
SOMATIC CELLS
TRACER TECHNIQUES
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550301* - Cytology- Tracer Techniques