Structural coupling of troponin C and actomyosin in muscle fibers.
EPR of spin labeled TnC at Cys98 was used to explore the possible structural coupling between TnC in the thin filament and myosin trapped in the intermediate states of ATPase cycle. Weakly attached myosin heads (trapped by low ionic strength, low temperature and ATP) did not induce structural changes in TnC as compared to relaxed muscle, as spin labeled TnC displayed the same narrow orientational distribution [Li, H.-C., and Fajer, P. G. (1994) Biochemistry 33, 14324]. Ca2+-binding alone resulted in disordering of the labeled domain of TnC. Additional conformational changes of TnC occurred upon the attachment of strongly bound, prepower stroke myosin heads (trapped by AlF4-). These changes were not present in ghost fibers which myosin had been removed, excluding direct effects of AlF4- on the orientation of TnC in muscle fibers. The postpower stroke heads (rigor.ADP/Ca2+ and rigor/Ca2+) induced further changes in the orientational distribution of labeled domain of TnC irrespective of the degree of cooperativity in thin filaments. We thus conclude that troponin C in thin filaments detects structural changes in myosin during force generation, implying that there is a structural coupling between actomyosin and TnC.
Li
H C
HC
Department of Biological Science and National High Magnetic Field Laboratory, Florida State University, Tallahassee 32306, USA.
Fajer
P G
PG
D000205:Actomyosin / Q000737:chemistry* / Q000502:physiology; D000244:Adenosine Diphosphate / Q000737:chemistry; D017607:Aluminum Compounds / Q000737:chemistry; D000818:Animals; D002118:Calcium / Q000737:chemistry; D004578:Electron Spin Resonance Spectroscopy; D005459:Fluorides / Q000737:chemistry; D066298:In Vitro Techniques; D009119:Muscle Contraction / Q000502:physiology; D018485:Muscle Fibers, Skeletal / Q000737:chemistry* / Q000502:physiology; D011487:Protein Conformation; D016658:Psoas Muscles; D011817:Rabbits; D013329:Structure-Activity Relationship; D019209:Troponin C / Q000737:chemistry* / Q000502:physiology
D017607:Aluminum Compounds; D019209:Troponin C; D000244:Adenosine Diphosphate; D000205:Actomyosin; D005459:Fluorides; D002118:Calcium; C032311:aluminum fluoride