The susceptibility of the glycoprotein from the purified (Na +, K +)-activated adenosine triphosphatase to tryptic and chymotryptic degradation with and without Na + and K +

Abstract

Purified (Na +, K +)-activated adenosine triphosphatase ((Na +, K +)-ATPase, ATP phosphohydrolase, EC 3.6.1.3) has been subjected to trypsin and chymotrypsin hydrolysis. The glycoprotein is much more resistant to proteolysis than the large chain. This differential susceptibility to proteolysis is not due to differences in the number of trypsin or chymotrypsin sensitive bonds because the two subunits are equally susceptible to proteolysis after isolation by preparative gel electrophoresis in sodium dodecyl sulfate. It is also not due to steric “shielding’ of the glycoprotein by the large chain or its proteolytic products: (1) The rate of digestion of the glycoprotein is not increased after 90% of the large chain is digested. (2) The majority of the large chain peptides are released into the supernatant upon degradation. It is concluded that the greater resistance of the glycoprotein to proteolysis is due to its native conformation. In the absence of the large chain, the susceptibility of the glycoprotein to tryptic degradation by K + and Na +. The evidence suggest that this decreased susceptibility was due to conformational changes in the glycoprotein. These specific ligand effects on proteolysis of the glycoprotein suggest that the glycoprotein may participate in Na + and K + binding by (Na +, K +)-ATPase.