Calorimetric studies of the thermal unfolding of smooth muscle myosin fragments and their complexes with ADP and phosphate analogs.
The thermal unfolding of turkey gizzard smooth muscle myosin subfragment 1 (S1) and heavy meromyosin (HMM) in the absence of added nucleotides, in the presence of ADP, and in S1 or HMM ternary complexes with ADP and Pi analogs, orthovanadate (Vi), beryllium fluoride (BeFx), or aluminum fluoride (AlF4-), have been studied by differential scanning calorimetry (DSC). It has been shown that the formation of these ternary complexes causes significant structural changes in S1 or in the heads of HMM which are reflected in a pronounced increase of the protein thermal stability. The effect of BeFx was less distinct than that of AlF4- or Vi. Phosphorylation of regulatory light chains (RLC) in S1 or in HMM had practically no influence on these effects. In general, the changes caused by various Pi analogs in smooth muscle S1 or HMM were similar to those observed earlier with skeletal muscle S1 devoid of RLC. It is concluded that RLC and their phosphorylation do not significantly affect the character of structural changes induced in motor domains of the HMM heads by the formation of ternary complexes HMM--ADP--Vi, HMM--ADP--AlF4-, and HMM--ADP--BeFx--stable analogs of the intermediate states of the HMM ATPase reaction, HMM.ADP.Pi and HMM. ATP.
Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, 119899, Russia. dpavlov@pcman.genebee.msu.su
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D000244:Adenosine Diphosphate / Q000378:metabolism*; D017607:Aluminum Compounds / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D001608:Beryllium / Q000378:metabolism / Q000494:pharmacology; D002152:Calorimetry, Differential Scanning; D005459:Fluorides / Q000378:metabolism / Q000494:pharmacology; D005895:Gizzard, Avian; D006358:Hot Temperature; D007700:Kinetics; D009130:Muscle, Smooth; D015879:Myosin Subfragments / Q000737:chemistry* / Q000378:metabolism*; D009218:Myosins / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D011489:Protein Denaturation; D017510:Protein Folding; D013816:Thermodynamics; D014422:Turkeys; D014638:Vanadates / Q000378:metabolism / Q000494:pharmacology
D017607:Aluminum Compounds; D015879:Myosin Subfragments; D010710:Phosphates; C050992:tetrafluoroaluminate; D014638:Vanadates; C035982:beryllium fluoride; D000244:Adenosine Diphosphate; D009218:Myosins; D001608:Beryllium; D005459:Fluorides