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Alterations in protein phosphorylation in human neutrophils (PMN) induced by a protein kinase C inhibitor, C-I

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:6279470
; ; ;

Protein phosphorylation is probably an important regulatory mechanism in PMN. The authors examined the effect of a novel protein kinase C (PKC) inhibitor, 1-(5-isoquinoline-sulfonyl) piperazine (C-I), on protein phosphorylation in human PMN. Purified PMN were loaded with TSP, pretreated with C-I for 5 min, and cell functions activated with phorbol myristate acetate or fMet-Leu-Phe for 30 sec. Cells were treated with protease inhibitor, sonicated, and centrifuged to yield cytosols and particulate fractions. Analysis of C-I-treated controls by SDS-PAGE and autoradiography revealed inhibition of background phosphorylation of essentially all resolved proteins, particularly prominent ones with molecular weights of 13.7, 43, 48, 60, 66, and 200 kd. Of these, the 48 kd protein has been correlated with activation of the respiratory burst in PMN. Fractions from cells stimulated with PMA or fMLP showed phosphorylation of at least 70-75 proteins. C-I pretreatment induced partial or complete inhibition of phosphorylation in virtually all proteins when cells were subsequently stimulated with PMA or fMLP. Thus, the authors data suggest 1) PKC may be active in some compartments of resting PMN, since C-I depressed resting levels of phosphorylation; 2) PKC plays a role in phosphorylation induced in PMN by activators such as PMA and fMLP, since all resolved proteins were affected by C-I treatment.

Research Organization:
Univ. Medical Center, Winston-Salem, NC
OSTI ID:
6279470
Report Number(s):
CONF-870644-
Journal Information:
Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 46:6; ISSN FEPRA
Country of Publication:
United States
Language:
English

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Related Subjects

550201 -- Biochemistry-- Tracer Techniques
560300* -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AMINO ACIDS
ANIMALS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL EFFECTS
BIOLOGICAL FUNCTIONS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOXYLIC ACIDS
CARCINOGENS
CHEMICAL REACTIONS
DAYS LIVING RADIOISOTOPES
ELECTROPHORESIS
ENZYME INHIBITORS
ENZYMES
ESTERS
FUNCTIONS
ISOTOPES
LEUKOCYTES
LIGHT NUCLEI
MAMMALS
MAN
MATERIALS
NEUTROPHILS
NUCLEI
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
PHENYLALANINE
PHORBOL ESTERS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHORYLATION
PHOSPHOTRANSFERASES
PRIMATES
PROTEINS
RADIOISOTOPES
TRANSFERASES
VERTEBRATES