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Local anesthetics have been reported to have several effects on platelets including:	1) inhibition of shape change, the release reaction and aggregation induced by several agents 2) dissolution of cytoskeletal structures and 3) activation of a protease activity which digests proteins of Mr 250,000 and 230,000. It was speculated that the last effect may be due to the Ca++-dependent protease previously described in platelets. I have recently observed that incubation of platelets with 1 mM dibucaine for brief periods of time (15-30 min) resulted in 50-100% enhancement of the initial slope of agglutination induced by either ristocetin or bovine von Willebrand factor (vWf) despite inhibiting the release reaction. If the platelets were formaldehyde-fixed after brief incubation, they also showed the enhancement (control 23 ± 3 U/min (mean ± SEM) and dibucaine 47 ± 6, p   1 hour, both ristocetin and bovine vWf-induced agglutination became progressively inhibited with almost total inhibition after several hours. This decrease in agglutination was associated with a progressive loss of a glycoprotein with the characteristics of GpIb (Mr 165,000 unreduced and 135,000 reduced) as judged by PAS-stained polyacrylamide gels. The presence of the platelet-impenetrable calcium chelator EDTA (10 mM) only partially prevented the decrease in agglutinability and loss of Gplb. I conclude that:	1. Short term incubation with dibucaine may improve the ristocetin cofactor assay. 2. A dibucaine-activated protease(s) appears to digest Gplb, probably accounting for the decreased agglutination with long incubations. 3. If operating from the outside of the platelet after leakage, at least some of the protease activity is not calcium dependent.

Dibucaine-Activated Platelet Protease(S) Digests GPIb-EDTA Only Partially Inhibits