title: Crenomytilus grayanus 40kDa calponin-like protein: cDNA cloning, sequence analysis, tissue expression, and post-translational modifications.
abstract: Calponin-like protein (CaP-40), a third major protein after actin and tropomyosin, has recently been identified by us in the Ca2+-regulated thin filaments of mussel Crenomytilus grayanus. It contains calponin homology domain, five calponin family repeats and possesses similar biochemical properties as vertebrate smooth muscle calponin. In this paper, we report a full-length cDNA sequence of CaP-40, study its expression pattern on mRNA and protein levels, evaluate CaP-40 post-translational modifications and perform protein-protein interaction analysis. The full-length sequence of CaP-40 consists of 398 amino acids and has high similarity to calponins among molluscan species. CaP-40 gene is widely expressed in mussel tissues, with the highest expression in adductor and mantle. Comparison of these data with protein content established by mass-spectrometry analysis revealed that the high mRNA content is mirrored by high protein levels for adductor smooth muscles. To provide unbiased insight into the function of CaP-40 and effect of its over-expression in adductor smooth muscle, we built protein-protein interaction network of identified Crenomytilus grayanus proteome. In addition, we showed that CaP-40 is subjected to post-translational N- and C-terminal acetylation at N127, G229 and G349 sites which potentially regulates its function in vivo.
authors.affiliation: A.V. Zhirmunsky Institute of Marine Biology, National Scientific Center of Marine Biology, Far East Branch of the Russian Academy of Sciences, Vladivostok, Russia; School of Biomedicine, Far Eastern Federal University, Vladivostok, Russia. Electronic address: osmatusovsky@gmail.com.
authors.forename: Oleg S
authors.initials: OS
authors.lastname: Matusovsky
authors.affiliation: A.V. Zhirmunsky Institute of Marine Biology, National Scientific Center of Marine Biology, Far East Branch of the Russian Academy of Sciences, Vladivostok, Russia.
authors.forename: Anna V
authors.initials: AV
authors.lastname: Dobrzhanskaya
authors.affiliation: A.V. Zhirmunsky Institute of Marine Biology, National Scientific Center of Marine Biology, Far East Branch of the Russian Academy of Sciences, Vladivostok, Russia; Far Eastern Federal University, Vladivostok, Russia.
authors.forename: Victoria V
authors.initials: VV
authors.lastname: Pankova
authors.affiliation: Laboratory of Biotechnology, Institute of Biology and Soil Science, Far East Branch of Russian Academy of Sciences, Vladivostok, Russia; Department of Biotechnology and Microbiology, The School of Natural Sciences, Far Eastern Federal University, Vladivostok, Russia.
authors.forename: Konstantin V
authors.initials: KV
authors.lastname: Kiselev
authors.affiliation: A.V. Zhirmunsky Institute of Marine Biology, National Scientific Center of Marine Biology, Far East Branch of the Russian Academy of Sciences, Vladivostok, Russia.
authors.forename: Ulyana V
authors.initials: UV
authors.lastname: Girich
authors.affiliation: A.V. Zhirmunsky Institute of Marine Biology, National Scientific Center of Marine Biology, Far East Branch of the Russian Academy of Sciences, Vladivostok, Russia.
authors.forename: Nikolay S
authors.initials: NS
authors.lastname: Shelud'ko
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D018076:DNA, Complementary; D008840:Microfilament Proteins / Q000235:genetics* / Q000378:metabolism*; D009130:Muscle, Smooth / Q000166:cytology / Q000378:metabolism*; D049879:Mytilidae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D010802:Phylogeny; D011487:Protein Conformation; D060066:Protein Interaction Maps; D011499:Protein Processing, Post-Translational; D017421:Sequence Analysis; D053719:Tandem Mass Spectrometry; D000096985:Calponins
chemical_list: D002135:Calcium-Binding Proteins; D018076:DNA, Complementary; D008840:Microfilament Proteins; D000096985:Calponins
keywords: Acetylation; Calponin-like protein; Mass-spectrometry; Post-translational modifications; Thin filaments