Human sperm protamines. Amino-acid sequences of two forms of protamine P2

Abstract

Human protamine P2 was purified to homogeneity by solubilizing whole spermatozoa in guanidinium · HCl containing 2‐mercaptoethanol, alkylating the resulting protamine thiols with vinylpyridine, removing acid‐insoluble material by acid dialysis and using CM‐cellulose chromatography to remove non‐protamine basic proteins and separate protamines P1 and P2. The P2 preparation contained two components, P2a and P2b, which were sequenced completely without being separated. The peptides obtained from thermolysin and endoproteinase Lys‐C digestions were purified by reverse‐phase high‐pressure liquid chromatography and sequenced using a gasphase sequencer. P2a contains 57 amino acids and has a relative molecular mass of 7636 while P2b contains 54 amino acids, which are identical to residues 4–57 of P2a, and has a relative molecular mass of 7242. Protamine P2a is approximately 50% homologous with human protamine P1. The amino acid sequence of P2a is: