Electron microscopic visualization of the SH1 thiol of myosin by the use of an avidin-biotin system.
One of the reactive thiols in the myosin head, SH1, was covalently labeled with a biotin derivative, N-iodoacetyl-N'-biotinylhexylenediamine. When 50% of the SH1 thiol was modified with the biotin reagent as judged from measurements of ATPase activities, the biotinylated myosin bound one mole of avidin per mole of myosin at the saturating level. The avidin-myosin complex was readily formed in the presence of MgADP or MgATP. Peptide maps of the biotinylated myosin revealed that SH1 is actually the site of biotinylation with N-iodoacetyl-N'-biotinylhexylenediamine. Electron microscopic examination of the avidin-myosin complex showed that the attachment site of avidin on the myosin head is 130 A from the head-rod junction, indicating that the SH1 thiol is located there.
D000595:Amino Acid Sequence; D000818:Animals; D001360:Avidin; D001710:Biotin / Q000031:analogs & derivatives; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D046911:Macromolecular Substances; D008301:Maleimides; D008854:Microscopy, Electron; D009218:Myosins; D011817:Rabbits; D013438:Sulfhydryl Compounds
D046911:Macromolecular Substances; D008301:Maleimides; D013438:Sulfhydryl Compounds; D001360:Avidin; C019347:N-(7-dimethylamino-4-methylcoumarinyl)maleimide; D001710:Biotin; C043329:N-iodoacetyl-N'-biotinylhexylenediamine; D009218:Myosins