Characterization of a type I regulatory subunit of cAMP-dependent protein kinase from the bivalve mollusk Mytilus galloprovincialis

Díaz-Enrich, María J.; Ibarguren, Izaskun; Hellman, Ulf; Villamarı́n, J.Antonio

doi:10.1016/s0003-9861(03)00259-5

Cited by 12 publications

(9 citation statements)

References 42 publications

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“…Work from 1972 to 1988 showed that agents that relaxed catch (e.g., serotonin) increased muscle cAMP levels, and that chemical treatments that increased cAMP relaxed catch {Achazi, 1979 36504 /id; Köhler, 1980 36506 / id;Achazi, 1974 35209 /id;Cole, 1972 449 /id;Gies, 1986 36300 /id;Gies, 1988 23435 / id;Marchand-Dumont, 1975 36296 /id;Matsuura, 1984 480 /id;Painter, 1982 54 /id;Pfitzer, 1982 23455 /id}. Subsequent work isolated and characterized the regulatory and catalytic subunits of the cAMP-dependent protein kinase (Cao et al, 1995a(Cao et al, ,b, 1996Rodríguez et al, 1998;Díaz-Enrich et al, 2003;Bardales et al, 2004;Béjar and Villamarín, 2006). The importance of protein phosphorylation was further emphasized by the observation that a calcineuron type phosphatase was required for catch (Castellani and Cohen, 1992).…”

Section: Unique Properties Due To Acto-myosin Interaction 1: Catchmentioning

confidence: 99%

Invertebrate muscles: Thin and thick filament structure; molecular basis of contraction and its regulation, catch and asynchronous muscle

Hooper

Hobbs

Thuma

2008

Progress in Neurobiology

131

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This is the second in a series of canonical reviews on invertebrate muscle. We cover here thin and thick filament structure, the molecular basis of force generation and its regulation, and two special properties of some invertebrate muscle, catch and asynchronous muscle. Invertebrate thin filaments resemble vertebrate thin filaments, although helix structure and tropomyosin arrangement show small differences. Invertebrate thick filaments, alternatively, are very different from vertebrate striated thick filaments and show great variation within invertebrates. Part of this diversity stems from variation in paramyosin content, which is greatly increased in very large diameter invertebrate thick filaments. Other of it arises from relatively small changes in filament backbone structure, which results in filaments with grossly similar myosin head placements (rotating crowns of heads every 14.5 nm) but large changes in detail (distances between heads in azimuthal registration varying from three to thousands of crowns). The lever arm basis of force generation is common to both vetebrates and invertebrates, and in some invertebrates this process is understood on the near atomic level. Invertebrate actomyosin is both thin (tropomyosin:troponin) and thick (primarily via direct Ca ++ binding to myosin) filament regulated, and most invertebrate muscles are dually regulated. These mechanisms are well understood on the molecular level, but the behavioral utility of dual regulation is less so. The phosphorylation state of the thick filament associated giant protein, twitchin, has been recently shown to be the molecular basis of catch. The molecular basis of the stretch activation underlying asynchronous muscle activity, however, remains unresolved.

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Section: Unique Properties Due To Acto-myosin Interaction 1: Catchmentioning

confidence: 99%

Invertebrate muscles: Thin and thick filament structure; molecular basis of contraction and its regulation, catch and asynchronous muscle

Hooper

Hobbs

Thuma

2008

Progress in Neurobiology

131

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“…In previous work we had been reported that R myt1 and R myt2 have specific structural and biochemical properties besides its common ability to inhibit the C-subunit in the absence of cAMP. First, only R myt2 , but not R myt1 , was phosphorylated in vitro by the own PKA C-subunit (Cao et al 1995;Díaz-Enrich et al 2003), and also by casein kinase-2 (CK2). CK2-mediated phosphorylation significantly decreases the ability of R myt2 to inhibit C-subunit in the absence of cAMP (Bardales et al 2007).…”

Section: Discussionmentioning

confidence: 99%

“…Polyclonal antisera against R myt1 and R myt2 were obtained in mouse and rabbit, respectively, as previously described (Cao et al 1995;Díaz-Enrich et al 2003). Antibodies were purified from rabbit sera or from mouse ascitic fluid through a Protein G Superose HR 10/2 chromatography in a FPLC system (GE Healthcare Bioscience, Uppsala, Sweden).…”

Section: Mantle Extractsmentioning

confidence: 99%

“…In order to better understand these regulatory mechanisms, we carried out a study to characterize the PKA from the sea mussel Mytilus galloprovincialis. To date, we have identified and purified two different isoforms of the R-subunit, named R myt1 and R myt2 , whose partial sequences were homologous to those of mammalian RI-and RII-type, respectively (Díaz Enrich et al 2003; Bardales et al 2007). Although both R myt1 and R myt2 show identical apparent molecular mass, they differ at their physico-chemical, biochemical and immunogenic properties (Cao et al 1995;Rodriguez et al 1998; Bardales et al 2007).…”

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confidence: 99%

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Differential distribution of cAMP-dependent protein kinase isoforms in the mantle of the bivalve mollusc Mytilus galloprovincialis

2009

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Two different isoforms of the regulatory (R) subunit of cAMP-dependent protein kinase (PKA), named R(myt1) and R(myt2), had previously been identified in the sea mussel Mytilus galloprovincialis. R(myt1) and R(myt2) were differentially distributed in the various cell types comprising the mussel mantle. R(myt1) was found the only isoform to be present in the auxiliary cells of female follicles and the cubic epithelium of the middle fold of mantle edge. In contrast, only the R(myt2) isoform was detected in the subepithelial connective tissue, the endothelium of haemolymph vessels, the adipogranular and vesicular cells of reserve connective tissue, and the spermatozoa. Finally, both R(myt1) and R(myt2) coexist in some cell types but they show a different cellular localization: R(myt1) was localized in the cytoplasm whereas R(myt2) was mainly detected in the cell apical edge, cell periphery, cilia and sperm flagella. Interestingly, both R(myt1) and R(myt2) were absent in oocytes within the female gonadal follicles but, in contrast, they were highly expressed in follicle-released oocytes collected after spawning. Taken together the results show that mussel PKA isoforms are differentially distributed in the mantle cell types, which suggests that they are involved in the regulation of distinct cellular functions. On the other hand, the expression of R(myt1) and R(myt2) proteins is associated with the meiosis resumption of oocytes at the prophase I stage, which occurs in parallel to spawning.

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“…No significant differences among C-subunit isoforms regarding the values of apparent K m for Kemptide and V max were observed. Furthermore, all the mussel isozymes were inhibited by the protein kinase inhibitor peptide [PKI (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)] with similar I 50 values ( Table 2). The ability of mussel C-subunit isoforms to phosphorylate proteins in vitro was also investigated.…”

Section: Kinetic Characterization Of C-subunit Isoforms and Protein Pmentioning

confidence: 99%

Identification of multiple isoforms of the cAMP‐dependent protein kinase catalytic subunit in the bivalve mollusc Mytilus galloprovincialis

2008

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Several isoforms of the cAMP‐dependent protein kinase catalytic subunit (C‐subunit) were separated from the posterior adductor muscle and the mantle tissues of the sea mussel Mytilus galloprovincialis by cation exchange chromatography, and identified by: (a) protein kinase activity; (b) antibody recognition; and (c) peptide mass fingerprinting. Some of the isozymes seemed to be tissue‐specific, and all them were phosphorylated at serine and threonine residues and showed slight but significant differences in their apparent molecular mass values, which ranged from 41.3 to 44.5 kDa. The results from the MS analysis suggest that at least some of the mussel C‐subunit isoforms arise as a result of alternative splicing events. Furthermore, several peptide sequences from mussel C‐subunits, determined by de novo sequencing, showed a high degree of homology with the mammalian Cα‐isoform, and contained some structural motifs that are essential for catalytic function. On the other hand, no significant differences were observed in the kinetic parameters of C‐subunit isoforms, determined by using synthetic peptides as substrate and inhibitor. However, the C‐subunit isoforms separated from the mantle tissue differed in their ability to phosphorylate in vitro some proteins present in a mantle extract.

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Characterization of a type I regulatory subunit of cAMP-dependent protein kinase from the bivalve mollusk Mytilus galloprovincialis

Cited by 12 publications

References 42 publications

Invertebrate muscles: Thin and thick filament structure; molecular basis of contraction and its regulation, catch and asynchronous muscle

Invertebrate muscles: Thin and thick filament structure; molecular basis of contraction and its regulation, catch and asynchronous muscle

Differential distribution of cAMP-dependent protein kinase isoforms in the mantle of the bivalve mollusc Mytilus galloprovincialis

Identification of multiple isoforms of the cAMP‐dependent protein kinase catalytic subunit in the bivalve mollusc Mytilus galloprovincialis

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