Inactivation, subunit dissociation, aggregation, and unfolding of myosin subfragment 1 during guanidine denaturation
| Autor: | Jean Jacques Bechet, Muriel Nozais, Maurice Houadjeto |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
Protein Denaturation
Circular dichroism Protein Conformation Hydrochloride Protein subunit ATPase Size-exclusion chromatography Guanidines Biochemistry Dissociation (chemistry) chemistry.chemical_compound Myosin Animals Chymotrypsin Guanidine Cation Transport Proteins Adenosine Triphosphatases biology Circular Dichroism Muscles Myosin Subfragments Kinetics chemistry biology.protein Biophysics Ca(2+) Mg(2+)-ATPase Rabbits Protein Binding |
| Zdroj: | Biochemistry. 31:1210-1215 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi00119a034 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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