title: Does Interaction between the Motor and Regulatory Domains of the Myosin Head Occur during ATPase Cycle? Evidence from Thermal Unfolding Studies on Myosin Subfragment 1.
abstract: Myosin head (myosin subfragment 1, S1) consists of two major structural domains, the motor (or catalytic) domain and the regulatory domain. Functioning of the myosin head as a molecular motor is believed to involve a rotation of the regulatory domain (lever arm) relative to the motor domain during the ATPase cycle. According to predictions, this rotation can be accompanied by an interaction between the motor domain and the C-terminus of the essential light chain (ELC) associated with the regulatory domain. To check this assumption, we applied differential scanning calorimetry (DSC) combined with temperature dependences of fluorescence to study changes in thermal unfolding and the domain structure of S1, which occur upon formation of the ternary complexes S1-ADP-AlF4- and S1-ADP-BeFx that mimic S1 ATPase intermediate states S1**-ADP-Pi and S1*-ATP, respectively. To identify the thermal transitions on the DSC profiles (i.e. to assign them to the structural domains of S1), we compared the DSC data with temperature-induced changes in fluorescence of either tryptophan residues, located only in the motor domain, or recombinant ELC mutants (light chain 1 isoform), which were first fluorescently labeled at different positions in their C-terminal half and then introduced into the S1 regulatory domain. We show that formation of the ternary complexes S1-ADP-AlF4- and S1-ADP-BeFx significantly stabilizes not only the motor domain, but also the regulatory domain of the S1 molecule implying interdomain interaction via ELC. This is consistent with the previously proposed concepts and also adds some new interesting details to the molecular mechanism of the myosin ATPase cycle.
authors.affiliation: A.N. Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, Russia; Department of Biochemistry, School of Biology, Moscow State University, Moscow, Russia; Department of Biotechnology, School of Biology, Vyatka State University, Kirov, Russia.
authors.forename: Daria S
authors.initials: DS
authors.lastname: Logvinova
authors.affiliation: A.N. Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Denis I
authors.initials: DI
authors.lastname: Markov
authors.affiliation: A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia.
authors.forename: Olga P
authors.initials: OP
authors.lastname: Nikolaeva
authors.affiliation: A.N. Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Nikolai N
authors.initials: NN
authors.lastname: Sluchanko
authors.affiliation: King's College London, London, United Kingdom.
authors.forename: Dmitry S
authors.initials: DS
authors.lastname: Ushakov
authors.affiliation: A.N. Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, Russia; A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia.
authors.forename: Dmitrii I
authors.initials: DI
authors.lastname: Levitsky
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D002152:Calorimetry, Differential Scanning; D005453:Fluorescence; D006801:Humans; D008958:Models, Molecular; D015879:Myosin Subfragments / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D058767:Protein Unfolding; D011817:Rabbits; D013696:Temperature; D014364:Tryptophan / Q000378:metabolism
chemical_list: D015879:Myosin Subfragments; D000244:Adenosine Diphosphate; D014364:Tryptophan; D000251:Adenosine Triphosphatases