Isolation of subunits from trypsin-cleaved sarcoplasmic reticulum Ca2+ transport adenosine triphosphatase.

Controlled tryptic digestion of purified rat skeletal muscle sarcoplasmic reticulum (Ca2+ + Mg2+)-adenosine triphosphate yields two products designated Fragments 3a and 3b with molecular weights of 65,000 and 56,000 respectively. The isolation of these products in high yield should facilitate exploration of the molecular characteristics of this adenosine triphosphatase. A simple, rapid method for accomplishing this isolation was developed which provides a high yield and utilizes mild conditions. The fragments obtained by this method were used to determine the phospholipid and sulfhydryl contents of Fragments 3a and 3b. In addition, information was obtained on the orientation of these adenosine triphosphatase components in the enzyme lipoprotein complex.