Immunocytochemical demonstration for caldesmon and actin in the striated and smooth muscle cells and non-muscular cells of various organs of rats.

Ban, Tadanobu; Ishimura, Kazunori; Fujita, Hisao; Sobue, Kenji; Kakiuchi, Shiro

doi:10.1267/ahc.17.331

Cited by 17 publications

(12 citation statements)

References 15 publications

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“…Caldesmon is broadly distributed among tissues and species (Kakiuchi et al, 1983;Ban et al, 1984;Owada et al, 1984;Bretscher & Lynch, 1985;Marston & Lehman, 1985;Ngai & Walsh, 1985b;Clark et al, 1986), including smooth, skeletal and cardiac muscles and a variety --of "non-muscle tissues. The demonstration of caldesmon in isolated rat cardiac myocytes confirms the presence of this protein in striated-muscle cells, since one can rule out possible contamination by vascular Table 3.…”

Section: Discussionmentioning

confidence: 99%

“…Actin and calmodulin compete for binding to caldesmon only in the presence of micromolar concentrations of Ca2+. Immunofluorescence studies (Ban et al, 1984;Owada et al, 1984;Bretscher & Lynch, 1985) and characterization of isolated smooth-muscle thin filaments (Marston & Lehman, 1985)-suggest that caldesmon is associated with actin filaments in vivo. The tissue content of caldesmon has been estimated to be 1 mol/42 actin monomers (Bretscher, 1984) or 11.1 LM (Ngai & Walsh, 1985a) in chicken gizzard and 1 mol/26 actin monomers in chicken gizzard, rabbit stomach, sheep aorta and sheep trachea (Marston & Lehman, 1985).…”

Section: Introductionmentioning

confidence: 99%

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The effects of caldesmon on the ATPase activities of rabbit skeletal-muscle myosin

Lim¹,

Walsh²

1986

Biochemical Journal

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We studied the effects of caldesmon, a major actin-and calmodulin-binding protein found in a variety of muscle and non-muscle tissues, on the various ATPase activities of skeletal-muscle myosin. Caldesmon inhibited the actin-activated myosin Mg2+-ATPase, and this inhibition was enhanced by tropomyosin. In the presence of the troponin complex and tropomyosin, caldesmon inhibited the Ca2+-dependent actomyosin Mg2+-ATPase; this inhibition could be partly overcome by Ca2+/calmodulin. Caldesmon, phosphorylated to the extent of 4 mol of PI/mol of caldesmon, inhibited the actin-activated myosin Mg2+-ATPase to the same extent as did non-phosphorylated caldesmon. Both inhibitions could be overcome by Ca2+/calmodulin. Caldesmon also inhibited the Mg2+-ATPase activity of skeletal-muscle myosin in the absence of actin; this inhibition also could be overcome by Ca2+/calmodulin. Caldesmon inhibited the Ca2+-ATPase activity of skeletal-muscle myosin in the presence or absence of actin, at both low (0.1 M-KCI) and high (0.3 M-KCI) ionic strength. Finally, caldesmon inhibited the skeletal-muscle myosin K+/EDTA-ATPase at 0.1 M-KCI, but not at 0.3 M-KCI. Addition of actin resulted in no inhibition of this ATPase by caldesmon at either 0.1 Mor 0.3 M-KCI. These observations suggest that caldesmon may function in the regulation of actin-myosin interactions in striated muscle and thereby modulate the contractile state of the muscle. The demonstration that caldesmon inhibits a variety of myosin ATPase activities in the absence of actin indicates a direct effect of caldesmon on myosin. The inhibition of the actin-activated Mg2+-ATPase activity of myosin (the physiological activity) may not be due therefore simply to the binding of caldesmon to the actin filament causing blockage of myosin-cross-bridge-actin interaction.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The effects of caldesmon on the ATPase activities of rabbit skeletal-muscle myosin

Lim¹,

Walsh²

1986

Biochemical Journal

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“…The smaller isoform of caldesmon, termed CD1, has been located in many non-muscle avian and mammalian tissues on the basis of its heat stability and crossreactivity with anti-h-caldesmon antibodies [42,43]. These tissues include platelets [44,45], fibroblasts [59], foetal smooth muscle [11,46], adrenal medulla [47], liver [48], erythrocytes [49], thyroid glands [50], absorptive epithelial cells [43], atherosclerotic plaques [10] and many cultured cell lines [46,51,63].…”

Section: Caldesmon Structurementioning

confidence: 99%

The molecular anatomy of caldesmon

Marston¹,

Redwood²

1991

Biochemical Journal

224

169

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“…The protein has been purified from several species, tissues, and cultured cells, e.g., chicken gizzard (Sobue et al 1981a;Bretscher 1984;, turkey gizzard (Malencik et al 1989), duck gizzard (Vorotnikov and Gusev 1991), bovine aorta (Clark et al 1986;Lash et al 1986), hog stomach (Fiirst et al 1986), human platelet (Dingus et al 1986), bovine adrenal medulla (Sobue et al 1985), bovine liver (Litchfield and Ball 1987), rabbit liver (Stafford et al 1990), molluscan striated muscle (Bartegi et al 1989), and REF4A cells (YamashiroMatsumura and Matsumura 1988). Caldesmon has been identified in many other tissues and cultured cells by immunocytochemical and Western blotting techniques (Ban et al 1984;Bretscher and Lynch 1985;Ngai and Walsh 1985a;Clark et al 1986). Immunofluorescence microscopy localized caldesmon to the stress fibers and ruffling membrane of cultured cells with a periodicity similar to the distribution of tropomyosin (Bretscher and Lynch 1985), consistent with its binding to actin and recovery in native thin filament preparations from smooth muscle (Marston and Lehman 1985;Ngai et al 1987).…”

Section: Caldesmonmentioning

confidence: 99%

Calcium-dependent mechanisms of regulation of smooth muscle contraction

Walsh¹

1991

Biochem. Cell Biol.

138

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The contractile state of smooth muscle is regulated primarily by the sarcoplasmic (cytosolic) free Ca2+ concentration. A variety of stimuli that induce smooth muscle contraction (e.g., membrane depolarization, alpha-adrenergic and muscarinic agonists) trigger an increase in sarcoplasmic free [Ca2+] from resting levels of 120-270 to 500-700 nM. At the elevated [Ca2+], Ca2+ binds to calmodulin, the ubiquitous and multifunctional Ca(2+)-binding protein. The interaction of Ca2+ with CaM induces a conformational change in the Ca(2+)-binding protein with exposure of a site(s) of interaction with target proteins, the most important of which in the context of smooth muscle contraction is the enzyme myosin light chain kinase. The interaction of calmodulin with myosin light chain kinase results in activation of the kinase that catalyzes phosphorylation of myosin at serine-19 of each of the two 20-kDa light chains (native myosin is a hexamer composed of two heavy chains (230 kDa each) and two pairs of light chains (one pair of 20 kDa each and the other pair of 17 kDa each)). This simple phosphorylation reaction triggers cycling of myosin cross-bridges along actin filaments and the development of force. Relaxation of the muscle follows removal of Ca2+ from the sarcoplasm, whereupon calmodulin dissociates from myosin light chain kinase regenerating the inactive kinase; myosin is dephosphorylated by myosin light chain phosphatase(s), whereupon it dissociates and remains detached from the actin filament and the muscle relaxes. A substantial body of evidence has been accumulated in support of this central role of myosin phosphorylation-dephosphorylation in the regulation of smooth muscle contraction. However, a wide range of physiological and biochemical studies supports the existence of additional, secondary Ca(2+)-dependent mechanisms that can modulate or fine-tune the contractile state of the smooth muscle cell. Three such mechanisms have emerged: (i) the actin-, tropomyosin-, and calmodulin-binding protein, calponin; (ii) the actin-, myosin-, tropomyosin-, and calmodulin-binding protein, caldesmon; and (iii) the Ca(2+)- and phospholipid-dependent protein kinase (protein kinase C).

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Immunocytochemical demonstration for caldesmon and actin in the striated and smooth muscle cells and non-muscular cells of various organs of rats.

Cited by 17 publications

References 15 publications

The effects of caldesmon on the ATPase activities of rabbit skeletal-muscle myosin

The effects of caldesmon on the ATPase activities of rabbit skeletal-muscle myosin

The molecular anatomy of caldesmon

Calcium-dependent mechanisms of regulation of smooth muscle contraction

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