Rapid Proteolysis of Brain MAP-1 Related Cytoskeleton-Associated 350kd Protein by Purified Calpain

Abstract

Microtubule associated protein-1 of brain and its intracellular 350kd analogues were highly sensitive to purified Ca²⁺-dependent cysteine proteinase (calpain). After 15 second digestion, we detected intermediate degradation products of MAP-1 by immunoblotting using anti-MAP-1 antibody as 290, 260, 220, 170, 140, 112, 80, 68, and 32kd polypeptides. These values corresponded to the molecular weights of the immunoreactive polypeptides of microtubule-enriched cytoskeletons isolated from HeLa and SV-3Y1 cells, suggesting the action of endogenous calpain on intracellular MAP-1 analogues in vivo or during the course of preparation.