Isolation and immunological characterization of the mammalian 32-/34-kDa stress protein.
Challenge of mammalian cells with heavy metals or sulfhydryl-reactive agents including sodium arsenite induces the de novo synthesis of a 32-/34-kDa stress protein (p32) (M. M. Caltabiano, T. P. Koestler, G. Poste, and R. G. Greig (1986) J. Biol. Chem. 261, 13,381). Here we report that antibody prepared against p32/p34 purified from human A375 melanoma cells immunoprecipitated an antigen of similar molecular mass from a panel of human, rat, and murine cells following challenge with sodium arsenite. No reactivity was observed in lysates from control, uninsulted cultures. The precise molecular mass of the arsenite-induced antigen was species-specific: 32 kDa (human and rat) and 34 kDa (murine). Indirect immunofluorescence analysis using affinity-purified monospecific IgG demonstrated that p32/p34 was localized to the cytoplasm and displayed a perinuclear distribution.
Caltabiano
M M
MM
Department of Cell Biology, Smith Kline & French Laboratories, Philadelphia, Pennsylvania 19101.
Poste
G
G
Greig
R G
RG
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000941:Antigens / Q000302:isolation & purification; D001151:Arsenic / Q000494:pharmacology*; D018053:Arsenites; D002460:Cell Line; D000939:Epitopes; D005455:Fluorescent Antibody Technique; D006360:Heat-Shock Proteins / Q000302:isolation & purification*; D006801:Humans; D007106:Immune Sera; D051379:Mice; D008970:Molecular Weight; D051381:Rats; D017670:Sodium Compounds; D013045:Species Specificity; D013312:Stress, Physiological / Q000209:etiology / Q000378:metabolism*
D000906:Antibodies; D000941:Antigens; D018053:Arsenites; D000939:Epitopes; D006360:Heat-Shock Proteins; D007106:Immune Sera; D017670:Sodium Compounds; C017947:sodium arsenite; D001151:Arsenic