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Activation of MAP kinases and phosphorylation of caldesmon in canine colonic smooth muscle.
Gerthoffer WT
,
Yamboliev IA
,
Shearer M
,
Pohl J
,
Haynes R
,
Dang S
,
Sato K
,
Sellers JR
.
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1. Phosphorylation of caldesmon was assayed in canine colonic circular smooth muscle strips labelled with 32P and stimulated with 10 microM acetylcholine. Caldesmon was isolated by two-dimensional non-equilibrium pH gel electrophoresis. Stimulation with acetylcholine increased caldesmon phosphorylation significantly from a basal level of 0.6 +/- 0.07 to 1.1 +/- 0.15 mol P1 (mol caldesmon)-1 after 2 min. 2. MAP kinase activities were measured in SDS extracts of muscle by a gel reconstitution method using myelin basic protein. Myelin basic protein kinase activities were observed at 38, 44, 50 and 57 kDa by the gel reconstitution method. Endogenous caldesmon kinase activities were also identified by the gel reconstitution method at 38, 44 and 50 kDa. The 38 and 44 kDa kinases comigrated with proteins labelled by anti-ERK1 MAP kinase antibodies on Western blots. Both 38 and 44 kDa MBP kinase activities increased significantly during contractions induced by 10 microM acetylcholine, 0.1 microM neurokinin A and 70 mM potassium. 3. Phorbol dibutyrate (0.1 microM) potentiated activation of MAP kinases and contraction of depolarized muscles while producing a decrease in fura-2 fluorescence ratio. This suggests that protein kinase C activation is coupled to MAP kinase activity in colonic smooth muscle. 4. MAP kinases isolated form muscle homogenates by Mono Q chromatography were assayed using the specific MAP kinase substrate peptide APRTPGGRR. Stimulation of muscles for 2 min with 10 microM acetylcholine activated both ERK1 and ERK2 MAP kinase activities 2-fold. 5. To determine the effects of caldesmon phosphorylation by MAP kinase on the cross-bridge cycle, actin sliding velocity was measured with an in vitro motility assay. Unphosphorylated turkey gizzard caldesmon (3 microM) significantly reduced mean sliding velocity. Phosphorylation of caldesmon with sea star ERK1 MAP kinase reversed the inhibitory effect of caldesmon on sliding velocity. The results are consistent with a protein kinase cascade being activated by contractile agonists in gastrointestinal smooth muscle which activates ERK MAP kinases leading to phosphorylation of caldesmon. Phosphorylation of caldesmon in vivo may reverse inhibitory influences of caldesmon on cross-bridge cycling.
Adam,
Activation of mitogen-activated protein kinase in porcine carotid arteries.
1995, Pubmed
Adam,
Activation of mitogen-activated protein kinase in porcine carotid arteries.
1995,
Pubmed
Adam,
Phosphorylation of caldesmon in arterial smooth muscle.
1989,
Pubmed
Adam,
Phosphorylation sequences in h-caldesmon from phorbol ester-stimulated canine aortas.
1992,
Pubmed
Adam,
Identification of mitogen-activated protein kinase phosphorylation sequences in mammalian h-Caldesmon.
1993,
Pubmed
Bretscher,
Smooth muscle caldesmon. Rapid purification and F-actin cross-linking properties.
1984,
Pubmed
Childs,
Phosphorylation of smooth muscle caldesmon by mitogen-activated protein (MAP) kinase and expression of MAP kinase in differentiated smooth muscle cells.
1992,
Pubmed
,
Echinobase
Clark,
Vascular smooth muscle caldesmon.
1986,
Pubmed
Erickson,
Identification by mass spectrometry of threonine 97 in bovine myelin basic protein as a specific phosphorylation site for mitogen-activated protein kinase.
1990,
Pubmed
Geahlen,
Detection of protein kinase activity in sodium dodecyl sulfate-polyacrylamide gels.
1986,
Pubmed
Gerthoffer,
Myosin phosphorylation and calcium in tonic and phasic contractions of colonic smooth muscle.
1991,
Pubmed
Gonzalez,
Heterogeneous expression of four MAP kinase isoforms in human tissues.
1992,
Pubmed
Haeberle,
The effects of smooth muscle caldesmon on actin filament motility.
1992,
Pubmed
Hathaway,
A radioimmunoblotting method for measuring myosin light chain phosphorylation levels in smooth muscle.
1985,
Pubmed
Homsher,
Factors affecting movement of F-actin filaments propelled by skeletal muscle heavy meromyosin.
1992,
Pubmed
Ikebe,
Binding of caldesmon to smooth muscle myosin.
1988,
Pubmed
Katayama,
Mode of caldesmon binding to smooth muscle thin filament: possible projection of the amino-terminal of caldesmon from native thin filament.
1995,
Pubmed
Kolch,
Protein kinase C alpha activates RAF-1 by direct phosphorylation.
1993,
Pubmed
Lee,
Role of nonselective cation current in muscarinic responses of canine colonic muscle.
1993,
Pubmed
Ludowyke,
Antigen-induced secretion of histamine and the phosphorylation of myosin by protein kinase C in rat basophilic leukemia cells.
1989,
Pubmed
Mabuchi,
Electron microscopic images suggest both ends of caldesmon interact with actin filaments.
1993,
Pubmed
Mak,
Phosphorylation of caldesmon by cdc2 kinase.
1991,
Pubmed
Mak,
Phosphorylation of caldesmon by p34cdc2 kinase. Identification of phosphorylation sites.
1991,
Pubmed
Moreland,
Regulation of a smooth muscle contraction: a hypothesis based on skinned fiber studies.
1991,
Pubmed
Ngai,
The effects of phosphorylation of smooth-muscle caldesmon.
1987,
Pubmed
O'Farrell,
High resolution two-dimensional electrophoresis of basic as well as acidic proteins.
1977,
Pubmed
Okagaki,
In vitro movement of actin filaments on gizzard smooth muscle myosin: requirement of phosphorylation of myosin light chain and effects of tropomyosin and caldesmon.
1991,
Pubmed
Park,
Carbachol-induced protein phosphorylation changes in bovine tracheal smooth muscle.
1986,
Pubmed
Sanders,
Ionic mechanisms of electrical rhythmicity in gastrointestinal smooth muscles.
1992,
Pubmed
Sato,
Sensitization of the contractile system of canine colonic smooth muscle by agonists and phorbol ester.
1994,
Pubmed
Scott-Woo,
Kinase activity associated with caldesmon is Ca2+/calmodulin-dependent kinase II.
1990,
Pubmed
Shirinsky,
Inhibition of the relative movement of actin and myosin by caldesmon and calponin.
1992,
Pubmed
Stoscheck,
Quantitation of protein.
1990,
Pubmed
Sutherland,
Phosphorylation of caldesmon prevents its interaction with smooth muscle myosin.
1989,
Pubmed
Tanaka,
Phosphorylation of high-Mr caldesmon by protein kinase C modulates the regulatory function of this protein on the interaction between actin and myosin.
1990,
Pubmed
Wawrzynow,
Identification of the site phosphorylated by casein kinase II in smooth muscle caldesmon.
1991,
Pubmed
Zhang,
Muscarinic receptors in canine colonic circular smooth muscle. II. Signal transduction pathways.
1991,
Pubmed
Zhang,
Protein kinase regulation of muscarinic receptor signalling in colonic smooth muscle.
1993,
Pubmed