The Superfast Human Extraocular Myosin Is Kinetically Distinct from the Fast Skeletal IIa, IIb, and IId Isoforms
| Autor: | Leslie A. Leinwand, John C. Deacon, Michael A. Geeves, Marieke J. Bloemink, Daniel I. Resnicow |
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| Rok vydání: | 2013 |
| Předmět: |
Gene isoform
Myosin macromolecular substances Biology Biochemistry Sarcomere Fluorescence Mice 03 medical and health sciences 0302 clinical medicine Cardiac Myosins Animals Humans Myocyte Protein Structure-Function Molecular Biology Actin Cell Line Transformed 030304 developmental biology 0303 health sciences Binding Sites Myosin Heavy Chains Homology Modeling Skeletal Muscle Myosins Cell Biology Recombinant Proteins Isoenzymes Kinetics Oculomotor Muscles Organ Specificity Enzymology Muscle MYH7 Sequence Alignment 030217 neurology & neurosurgery Signal Transduction |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m113.488130 |
| Popis: | Background: Characterization of individual muscle myosin isoforms has been limited by the availability of pure samples of isoforms. Results: Human myosin isoforms expressed in mouse cell line all have different ADP affinities. Conclusion: The superfast extraocular myosin is kinetically distinct from the fast skeletal IIa, IId, and IIb isoforms. Significance: This is the first time adult human sarcomeric myosins have been isolated and characterized by transient kinetics. Humans express five distinct myosin isoforms in the sarcomeres of adult striated muscle (fast IIa, IId, the slow/cardiac isoform I/β, the cardiac specific isoform α, and the specialized extraocular muscle isoform). An additional isoform, IIb, is present in the genome but is not normally expressed in healthy human muscles. Muscle fibers expressing each isoform have distinct characteristics including shortening velocity. Defining the properties of the isoforms in detail has been limited by the availability of pure samples of the individual proteins. Here we study purified recombinant human myosin motor domains expressed in mouse C2C12 muscle cells. The results of kinetic analysis show that among the closely related adult skeletal isoforms, the affinity of ADP for actin·myosin (KAD) is the characteristic that most readily distinguishes the isoforms. The three fast muscle myosins have KAD values of 118, 80, and 55 μm for IId, IIa, and IIb, respectively, which follows the speed in motility assays from fastest to slowest. Extraocular muscle is unusually fast with a far weaker KAD = 352 μm. Sequence comparisons and homology modeling of the structures identify a few key areas of sequence that may define the differences between the isoforms, including a region of the upper 50-kDa domain important in signaling between the nucleotide pocket and the actin-binding site. |
| Databáze: | OpenAIRE |
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