A fluorescence temperature-jump study of conformational transitions in myosin subfragment 1.
The tryptophan fluorescence of unmodified myosin subfragment 1 (S1) from rabbit and chicken skeletal muscle with various nucleotides and phosphate analogues bound was measured after rapid temperature jumps. The fluorescence decreased during the temperature rise. Under some conditions, this decrease was followed by an increase, reflecting structural transitions within the protein. With adenosine 5'-[beta,gamma-imido]triphosphate (p[NH]ppA) or with ADP and BeF(x) bound, this rise was very rapid (reciprocal time constant approx. 2000 s(-1)) and varied only slightly with starting temperature, suggesting that, with these ligands, two different protein conformations were present in rapid equilibrium over a large temperature range. In the presence of ATP, the transient included several relaxation processes. Overall, the results suggest that complexes of S1 with ATP or with a number of other ligands exist as a mixture of two forms in temperature-dependent equilibrium. The results throw light on the finding of different forms of S1 in recent crystallographic studies and indicate a surprising lack of strong coupling between myosin's structural state and the nature of the nucleotide bound.
Urbanke
C
C
Medizinische Hochschule Hannover, Carl-Neuberg-Str. 1, D-30623 Hannover, Germany.
Wray
J
J
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001608:Beryllium / Q000494:pharmacology; D002645:Chickens; D004023:Dictyostelium / Q000378:metabolism; D005459:Fluorides / Q000494:pharmacology; D007700:Kinetics; D008024:Ligands; D008956:Models, Chemical; D018482:Muscle, Skeletal / Q000737:chemistry; D015879:Myosin Subfragments / Q000737:chemistry*; D009218:Myosins / Q000737:chemistry; D011487:Protein Conformation; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013816:Thermodynamics; D013997:Time Factors
D008024:Ligands; D015879:Myosin Subfragments; C035982:beryllium fluoride; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D009218:Myosins; D001608:Beryllium; D005459:Fluorides