Open AccessSelective Carbethoxylation of the Histidine Residues of Actin by Diethylpyrocarbonate
Author(s) -
Hegyi György,
Premecz György,
Sain Béla,
Mühlrád András
Publication year - 1974
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1974.tb03452.x
Subject(s) - histidine , chemistry , polymerization , actin , residue (chemistry) , monomer , ultracentrifuge , biochemistry , hydroxylamine , peptide , organic chemistry , amino acid , polymer
Histidine residues of actin were carbethoxylated with diethylpyrocarbonate. Four and three histidine residues per actin monomer were rapidly carbethoxylated in the G and F form, respectively. After partial Carbethoxylation of G‐actin a non‐polymerizable fraction could be separated by ultracentrifugation. The analysis of these fractions showed that one of the four fast‐reacting histidines of G‐actin plays an essential role in polymerization. Removal of the carbethoxy groups from the histidine residues by hydroxylamine treatment restored the ability to polymerize. The specificity of the reaction was tested by the use of [ 14 C]diethylpyrocarbonate. After tryptic digestion of non‐polymerizable carbethoxylated actin a single histidyl peptide was isolated by gel filtration and diagonal electrophoresis. The histidine residue essential for polymerization was identified as histidine‐40.