Inactivation, subunit dissociation, aggregation, and unfolding of myosin subfragment 1 during guanidine denaturation.
The effect of guanidine hydrochloride on ATPase activity, gel filtration, turbidity, exposure of thiol groups, far-UV circular dichroism, and the fluorescence emission intensity of myosin subfragment 1 (S-1) was studied under equilibrium conditions. It was found that the denaturation process involves several intermediate states. The enzymatic activity of S-1 is at first lost at very low concentrations of GdnHCl (lower than 0.5 M). At a slightly higher GdnHCl concentration (about 0.5 M), the light chains dissociate and this dissociation is closely followed by the formation of aggregates between the naked heavy chains of S-1 molecules in the guanidine hydrochloride range of concentrations 0.5-1 M. At GdnHCl concentrations above 1 M, aggregates gradually disappear and S-1 loses its secondary and tertiary structures. These phenomena are partly reversible, and ATPase activity is only partially recovered under highly limited conditions. These results are discussed in relation to the nature of myosin subunit assembly. The head fragment of 20 kDa is thus suggested to be implicated in the binding of light chain to heavy chain and in the self-association of free heavy chains.
Nozais
M
M
Laboratoire de Biologie Physicochimique, Unité Associée au CNRS 1131, Orsay, France.
Béchet
J J
JJ
Houadjeto
M
M
D000251:Adenosine Triphosphatases / Q000737:chemistry; D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase / Q000737:chemistry; D027682:Cation Transport Proteins; D002918:Chymotrypsin; D002942:Circular Dichroism; D019791:Guanidine; D006146:Guanidines / Q000494:pharmacology*; D007700:Kinetics; D009132:Muscles / Q000737:chemistry / Q000201:enzymology; D015879:Myosin Subfragments / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation; D011817:Rabbits
D027682:Cation Transport Proteins; D006146:Guanidines; D015879:Myosin Subfragments; D002918:Chymotrypsin; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase; C022761:potassium transporting ATPase; D019791:Guanidine