title: Kinetic mechanism of non-muscle myosin IIB: functional adaptations for tension generation and maintenance.
abstract: Besides driving contraction of various types of muscle tissue, conventional (class II) myosins serve essential cellular functions and are ubiquitously expressed in eukaryotic cells. Three different isoforms in the human myosin complement have been identified as non-muscle class II myosins. Here we report the kinetic characterization of a human non-muscle myosin IIB subfragment-1 construct produced in the baculovirus expression system. Transient kinetic data show that most steps of the actomyosin ATPase cycle are slowed down compared with other class II myosins. The ADP affinity of subfragment-1 is unusually high even in the presence of actin filaments, and the rate of ADP release is close to the steady-state ATPase rate. Thus, non-muscle myosin IIB subfragment-1 spends a significantly higher proportion of its kinetic cycle strongly attached to actin than do the muscle myosins. This feature is even more pronounced at slightly elevated ADP levels, and it may be important in carrying out the cellular functions of this isoform working in small filamentous assemblies.
authors.affiliation: Laboratory of Molecular Cardiology, NHLBI, National Institutes of Health, Bethesda, Maryland 20892-1762, USA.
authors.forename: Fei
authors.initials: F
authors.lastname: Wang
authors.forename: Mihaly
authors.initials: M
authors.lastname: Kovacs
authors.forename: Aihua
authors.initials: A
authors.lastname: Hu
authors.forename: John
authors.initials: J
authors.lastname: Limouze
authors.forename: Estelle V
authors.initials: EV
authors.lastname: Harvey
authors.forename: James R
authors.initials: JR
authors.lastname: Sellers
mesh_terms: D000199:Actins / Q000737:chemistry; D000205:Actomyosin / Q000737:chemistry; D000244:Adenosine Diphosphate / Q000737:chemistry; D000251:Adenosine Triphosphatases / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D008956:Models, Chemical; D024501:Nonmuscle Myosin Type IIB / Q000737:chemistry*; D010766:Phosphorylation; D020033:Protein Isoforms; D011721:Pyrenes / Q000737:chemistry; D011817:Rabbits; D013816:Thermodynamics; D013997:Time Factors
chemical_list: D000199:Actins; D018076:DNA, Complementary; D020033:Protein Isoforms; D011721:Pyrenes; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000205:Actomyosin; C030984:pyrene; D000251:Adenosine Triphosphatases; D024501:Nonmuscle Myosin Type IIB