Interference with myosin subfragment-1 binding by site-directed mutagenesis of actin.
Three N-terminal double mutants of beta-actin expressed in the yeast Saccharomyces cerevisiae have been characterized with respect to DNase-I interaction, N-terminal post-translational modification, polymerizability and myosin subfragment-1 binding. The results strongly support earlier suggestions that the acidic residues at the N-terminus of actin are part of the myosin-binding site, while they seem to be of no importance for the other aspects of actin biochemistry tested. The suitability of this expression system for production of recombinant actin in general is discussed.
Aspenström
P
P
Department of Developmental Biology, Uppsala University, Sweden.
Karlsson
R
R
D000199:Actins / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002645:Chickens; D002850:Chromatography, Gel; D003850:Deoxyribonuclease I / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D015879:Myosin Subfragments / Q000378:metabolism* / Q000648:ultrastructure; D011499:Protein Processing, Post-Translational; D012441:Saccharomyces cerevisiae / Q000235:genetics
D000199:Actins; D015879:Myosin Subfragments; D003850:Deoxyribonuclease I