Mapping of functional domains in the plasma membrane Ca2+ pump using trypsin proteolysis

Abstract

The purified erythrocyte Ca2+ pump has been exposed to trypsin under conditions designed to enrich the fragments of molecular mass 90, 85, 81, and 76 kDa, respectively. In SDS-polyacrylamide gels, these fragments are accompanied by a product of molecular mass about 33 kDa. N- and C-terminal sequencing of the fragments blotted on PVDF membranes has located the four high molecular mass fragments and the 33-kDa fragment within the pump structure. The work has extended previous work on the organization of the calmodulin-interacting domain of the pump (Zurini et al., 1984; Benaim et al., 1984) and has tentatively placed the domain of the pump which interacts with acidic phospholipids between transmembrane helices 2 and 3.