Procalpain I in cytoplasm is translocated onto plasma and granule membranes during platelet stimulation with thrombin and then activated on the membranes.
Thrombin stimulation of platelets resulted in changes in the subcellular localization of calpain I, with a concomitant alteration of its molecular weight as measured by immunoblotting. Calpain I in resting platelets was distributed as procalpain I, an 80 kDa form which does not exhibit the enzyme activity, and 83% of the total antigen was localized in the cytosol fraction. When platelets were stimulated with thrombin, the total content of calpain I antigen was not significantly changed as compared with that of the resting platelets, though a decrease in the cytosolic distribution of 80 kDa form (from 83% to 47% of the total antigen) was observed with concomitant appearance of the active 76 kDa and intermediate 78 kDa forms of calpain I and increase in the 80 kDa form in the granule and membrane fractions. These results indicated that calpain I was translocated from the cytosol to both the plasma and granule membranes as procalpain I and then activated on the membranes during platelet stimulation with thrombin.
Kuboki
M
M
Department of Clinical Biochemistry, Faculty of Pharmaceutical Sciences, Teikyo University, Kanagawa, Japan.
Ishii
H
H
Horie
S
S
Kazama
M
M
D001792:Blood Platelets / Q000187:drug effects / Q000201:enzymology*; D002154:Calpain / Q000097:blood* / Q000302:isolation & purification; D003594:Cytoplasmic Granules / Q000201:enzymology*; D003600:Cytosol / Q000201:enzymology; D004792:Enzyme Precursors / Q000097:blood* / Q000302:isolation & purification; D006801:Humans; D007425:Intracellular Membranes / Q000201:enzymology; D007770:L-Lactate Dehydrogenase / Q000097:blood / Q000302:isolation & purification; D008970:Molecular Weight; D013917:Thrombin / Q000494:pharmacology*
D004792:Enzyme Precursors; D007770:L-Lactate Dehydrogenase; D013917:Thrombin; D002154:Calpain; C098484:mu-calpain proenzyme