Partial purification and characterization of a protein lysine methyltransferase from plasmodia of Physarum polycephalum

Publisher Website
Google Scholar
Abstract
Plasmodia of P. polycephalum have an active protein lysine methyltransferase (S-adenosylmethionine:protein-lysine methyltransferase, EC 2.1.1.43). This enzyme was purified 40-fold with a 13% yield, and it catalyzes the transfer of methyl groups from S-adenosyl-L-methionine to the .epsilon.-amino group of lysine residues with formation of N.epsilon.-mono-, N.epsilon.-di-, and N.epsilon.-trimethyllysines in a molar ratio of 4:1:1 based on [14C]methyl incorporation into the methylated lysines. The ratio remains unchanged at all stages of the partial purification, as well as after fractionation by sucrose density gradient centrifugation and gel electrophoresis. The rate of protein methylation is time dependent, enzyme concentration dependent, and requires the presence of a sulfhydryl reducing agent for optimal activity. The enzyme has optimal activity at pH 8 and is inhibited by S-adenosyl-L-homocysteine and EDTA. Lysine-rich and arginine-rich histones serve as the most effective exogenous protein acceptors; P. polycephalum actomyosin is inactive, and chick skeletal myofibrillar proteins are 25% as effective as exogeneous mixed histones as substrates. Lysine, polylysine, RNase A, cytochrome c and bovine serum albumin are not methylated.