Solution structure of myosin-ADP-MgFn ternary complex by fluorescent probes and small-angle synchrotron X-ray scattering.
In the presence of excess amounts of fluorine, a physiological divalent cation, magnesium (Mg(2+)), forms a novel phosphate analogue, magnesium fluoride (MgFn). Park et al. [Biochim. Biophys. Acta 1430, 127-140 (1999)] previously demonstrated that MgADP. MgFn forms a complex with myosin subfragment-1 (S-1), and the S-1.ADP. MgFn ternary complex mimics a transient state in the activity cycle of ATPase. In the present study, localized conformations in the regions of highly reactive cysteine and lysine residues, Cys 707 (SH1), Cys 697 (SH2), and Lys 83 (RLR), which change their conformations markedly during ATP hydrolysis, were studied using fluorescent probes and chemical modification. The global shape of the complex was also studied using small angle X-ray solution scattering and compared it with other previously reported myosin.ADP. fluorometal ternary complexes. The results suggest that the overall conformation and localized functional regions of the complex are quite similar to those in the presence of ATP, indicating that the complex mimics the M(**).ADP.P(i) steady state.
Maruta
S
S
Department of Bioengineering, Faculty of Engineering, Soka University, Hachioji, Tokyo 192-8577, Japan. shinsaku@t.soka.ac.jp
Aihara
T
T
Uyehara
Y
Y
Homma
K
K
Sugimoto
Y
Y
Wakabayashi
K
K
D000244:Adenosine Diphosphate / Q000737:chemistry* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D000817:Anilino Naphthalenesulfonates / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002645:Chickens; D005456:Fluorescent Dyes / Q000378:metabolism; D005459:Fluorides / Q000737:chemistry* / Q000378:metabolism; D017616:Magnesium Compounds / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D018482:Muscle, Skeletal / Q000737:chemistry; D015879:Myosin Subfragments / Q000737:chemistry* / Q000378:metabolism*; D010069:Oxadiazoles / Q000737:chemistry / Q000378:metabolism; D018583:Pliability; D011485:Protein Binding; D011487:Protein Conformation; D012542:Scattering, Radiation; D012996:Solutions; D013050:Spectrometry, Fluorescence; D017356:Synchrotrons; D014302:Trinitrobenzenesulfonic Acid / Q000737:chemistry / Q000378:metabolism; D014965:X-Rays
D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; D017616:Magnesium Compounds; D015879:Myosin Subfragments; D010069:Oxadiazoles; D012996:Solutions; C031288:magnesium fluoride; D000244:Adenosine Diphosphate; C021310:2-(4'-maleimidylanilino)naphthalene-6-sulfonic acid; D000255:Adenosine Triphosphate; D014302:Trinitrobenzenesulfonic Acid; C046364:4-(aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole; D005459:Fluorides