Selective cleavage at lysine of the 50 kDa-20 kDa connector loop segment of skeletal myosin S-1 by endoproteinase Arg-C.
The reaction of endoproteinase Arg-C on the skeletal myosin head heavy chain was investigated through characterization of peptides and amino acid sequence analysis. The protease splits exclusively the 50 kDa-20 kDa junction at the lysine cluster spanning residues 639-641 and does not affect any other protease-sensitive region of the entire myosin heavy chain. The sensitivity of the cleavage to actin and nucleotide binding makes this protease a very specific conformational probe of S-1. The nicked S-1 derivative, containing an intact NH2-terminal 75 kDa fragment, may serve as a tool for gaining further insights into the domain structure and function of the myosin head.
Bertrand
R
R
Centre de Recherches de Biochimie Macromoléculaire du CNRS, INSERM U249, Université de Montpellier, France.
Derancourt
J
J
Kassab
R
R
D000199:Actins / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase / Q000378:metabolism; D003432:Cross-Linking Reagents; D010450:Endopeptidases / Q000378:metabolism*; D005456:Fluorescent Dyes; D008239:Lysine; D008970:Molecular Weight; D015879:Myosin Subfragments; D009218:Myosins / Q000378:metabolism*; D009282:Naphthalenesulfonates; D010446:Peptide Fragments / Q000378:metabolism*; D011487:Protein Conformation; D011817:Rabbits; D012697:Serine Endopeptidases; D014357:Trypsin / Q000378:metabolism
D000199:Actins; D003432:Cross-Linking Reagents; D005456:Fluorescent Dyes; D015879:Myosin Subfragments; D009282:Naphthalenesulfonates; D010446:Peptide Fragments; C006421:1,5-I-AEDANS; D000255:Adenosine Triphosphate; D010450:Endopeptidases; C044499:submandibular proteinase A; D012697:Serine Endopeptidases; D014357:Trypsin; D017301:Ca(2+) Mg(2+)-ATPase; D009218:Myosins; D008239:Lysine