Effect of ionic strength on the conformation of myosin subfragment 1-nucleotide complexes.
The effect of ionic strength on the conformation and stability of S1 and S1-nucleotide-phosphate analog complexes in solution was studied. It was found that increasing concentration of KCl enhances the reactivity of Cys(707) (SH1 thiol) and Lys(84) (reactive lysyl residue) and the nucleotide-induced tryptophan fluorescence increment. In contrast, high KCl concentration lowers the structural differences between the intermediate states of ATP hydrolysis in the vicinity of Cys(707), Trp(510) and the active site, possibly by increasing the flexibility of the molecule. High concentrations of neutral salts inhibit both the formation and the dissociation of the M**.ADP.Pi analog S1.ADP.Vi complex. High ionic strength profoundly affects the structure of the stable S1.ADP.BeF(x) complex, by destabilizing the M*.ATP intermediate, which is the predominant form of the complex at low ionic strength, and shifting the equilibrium to favor the M**.ADP.Pi state. The M*.ATP intermediate is destabilized by perturbation of ionic interactions possibly by disruption of salt bridges. Two salt-bridge pairs, Glu(501)-Lys(505) in the Switch II helix and Glu(776)-Lys(84) connecting the catalytic domain to the lever arm, seem most appropriate to consider for participating in the ionic strength-induced transition of the open M*.ATP to the closed M**.ADP.Pi state of S1.
Peyser
Y M
YM
Hebrew University Hadassah School of Dental Medicine, Institute of Dental Sciences, Department of Oral Biology, Jerusalem 91120, Israel.
Ajtai
K
K
Burghardt
T P
TP
Muhlrad
A
A
D000244:Adenosine Diphosphate / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000478:Alkylation; D000818:Animals; D002942:Circular Dichroism; D003545:Cysteine / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D018482:Muscle, Skeletal; D015879:Myosin Subfragments / Q000737:chemistry* / Q000378:metabolism*; D009711:Nucleotides / Q000378:metabolism*; D009994:Osmolar Concentration; D011188:Potassium / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D011487:Protein Conformation / Q000187:drug effects; D011817:Rabbits; D013997:Time Factors; D014301:Trinitrobenzenes / Q000378:metabolism; D014364:Tryptophan / Q000378:metabolism; D014638:Vanadates / Q000494:pharmacology
D015879:Myosin Subfragments; D009711:Nucleotides; D014301:Trinitrobenzenes; D014638:Vanadates; D000244:Adenosine Diphosphate; D014364:Tryptophan; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D008239:Lysine; D003545:Cysteine; D011188:Potassium