Updated Protocols and Comments on the Purification without Use of Activating Ligands of the Coupling Proteins N_s and N_i of the Hormone Sensitive Adenylyl Cyclase

Abstract

N_s and N_i have been purified without using NaF and Mg as stabilizing agents (Codina, J., Hildebrandt, J.D., Sekura, R.D., Birnbaumer, M., Bryan, J., Manclark, C.R. and Birnbaumer, L. [1984] J. Biol. Chem. 259, in press). Since the submission of that report, several modifications have been introduced to the purification procedure and additional fractions have been processed from which N proteins are obtained. This article describes the updated protocols and presents methodological details not included in the previous publication. The final products are N_s, the stimulatory N, N_i the inhibitory N, both of subunit structure αβγ, and a M_r=40,000 protein of βγ composition. They are obtained from human erythrocytes.