MECHANISM OF HISTAMINE RELEASE BY ALPHA-CHYMOTRYPSIN FROM ISOLATED RAT MAST CELLS
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Alpha-chymotrypsin (CT) was modified chemically and physically by the treatments with diisopropyl fluorophosphatc, L-(1-tosylamide-2-phenyl) ethylchloromethylketone, hydrogen peroxide and heat. After these treatments, CT lost or decreased both the enzymic activity and ability of releasing histamine from rat mast cells. Ca<SUP>++</SUP> was essential for histamine release by CT, while it enhanced only slightly the enzymic activity. Process of histamine release by CT could be separated into two stages: CT-dependent but not Ca<SUP>++</SUP>-dependent, and Ca<SUP>++</SUP>-dependent but not CT-dependent. The activated state of mast cells produced by CT decayed rapidly at 37°C in the absence of Ca<SUP>++</SUP>, but these cells responded to Ca<SUP>++</SUP> by adding CT once again, suggesting reconstitution of cell membrane structure affected by CT. Isoproterenol, epinephrine, prostaglandin E, , and dibutyryl-cyclic AMP (0.01-0.1 mM) did not inhibit release of histamine induced by CT. Neither theophylline (0.01-0.1 mM) alone nor the combinations of these cyclic AMP-active agents with theophylline inhibited the release of histamine. But, in the presence of papaverine (0.01-0.1 mM) a marked, dose-dependent inhibition was observed. These data suggest that 1) release of histamine by CT from rat mast cells is causally related to its hydrolytic activity, 2) this activity causes a reversible change on mast cell membrane which probably facilitates Ca<SUP>++</SUP>-influx through the cell membrane, and 3) there are subtle differences among CT, compound 48/80 and antigens concerning the effect of cyclic AMP-active agents in histamine-releasing mechanisms in mast cells.
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