scholarly journals T-tubule membranes from chicken skeletal muscle possess an enzymic cascade for degradation of extracellular ATP

1997 ◽  
Vol 327 (3) ◽  
pp. 899-907 ◽  
Author(s):  
Jerónimo DELGADO ◽  
Gloria MORO ◽  
Ana SABORIDO ◽  
Alicia MEGÍAS
Keyword(s):  
Skeletal Muscle ◽  
Concanavalin A ◽  
Hydrolytic Activity ◽  
Extracellular Atp ◽  
Purinergic Signalling ◽  
Activity Effect ◽  
Characterization Studies ◽  
Extracellular Side ◽  
Triton X ◽  
Chicken Skeletal Muscle

The chicken T-tubule Mg2+-ATPase is an integral membrane glycoprotein that presents properties different from those of other ATPases located in skeletal muscle cells and exhibits ATP-hydrolysing activity on the extracellular side of the transverse tubule (TT) membranes. In this study we demonstrate that TT vesicles purified from chicken skeletal muscle possess ecto-ADPase and ecto-5ʹ-nucleotidase activities that, along with ecto-ATPase, are able to sequentially degrade extracellular ATP to ADP, AMP and adenosine. Characterization studies of these TT ectonucleotidases revealed remarkable differences between ecto-ATPase and ecto-ADPase activities with respect to thermal stability, temperature dependence of the hydrolytic activity, effect of ionic strength, kinetic behaviour, divalent cation preference and responses to azide, N-ethylmaleimide, NaSCN, Triton X-100 and concanavalin A. Ecto-ATPase, but not ecto-ADPase, was inhibited by a polyclonal antibody against the chicken TT ecto-ATPase. On the basis of these results we propose that ATP and ADP hydrolysis are accomplished by two distinct enzymes and therefore the TT ecto-ATPase is not an apyrase. 5ʹ-Nucleotidase activity was inhibited by adenosine 5ʹ-[α,β-methylene]diphosphate and concanavalin A, followed simple Michaelis-Menten kinetics and was released from the membranes by treatment with phosphatidylinositol-specific phospholipase C, indicating that AMP hydrolysis in T-tubules is catalysed by a typical ecto-5ʹ-nucleotidase. Results obtained from electrophoresis experiments under native conditions suggest that ecto-ATPase, ecto-ADPase and 5ʹ-nucleotidase might be associated, forming functional complexes in the T-tubule membranes. The TT ectonucleotidases constitute an enzymic cascade for the degradation of extracellular ATP that might be involved in the regulation of purinergic signalling in the muscle fibre.

Regulation of transverse tubule ecto-ATPase activity in chicken skeletal muscle

2001 ◽  
Vol 353 (3) ◽  
pp. 521-529 ◽  
Author(s):  
Alicia MEGÍAS ◽  
M. Mar MARTÍNEZ-SENAC ◽  
Jerónimo DELGADO ◽  
Ana SABORIDO
Keyword(s):  
Skeletal Muscle ◽  
Atpase Activity ◽  
Cholesterol Oxidase ◽  
Cross Linking ◽  
Thermal Variation ◽  
Con A ◽  
Transverse Tubule ◽  
Membrane Components ◽  
Triton X ◽  
Chicken Skeletal Muscle

Transverse tubule (T-tubule) ecto-ATPase from chicken skeletal muscle is an integral membrane glycoprotein that seems to exist as a homodimer and exhibits unusual properties. Treatment of T-tubule membranes with concanavalin A (Con A) did not significantly affect the thermal variation of the fluorescence anisotropy of vesicles labelled with 1,6-diphenyl-1,3,5-hexatriene or trimethylammonium-1,6-diphenyl-1,3,5-hexatriene. Cross-linking of membrane components with glutaraldehyde elicited effects on ecto-ATPase activity very similar to those of Con A treatment: a severalfold increase in activity, a decrease in Triton X-100 sensitivity and a requirement to be present before ATP to exert its action. In addition, glutaraldehyde and Con A normalized the temperature dependence and the kinetic behaviour of the enzyme. Membrane-perturbing agents (detergents, alcohols and cholesterol oxidase), with the sole exception of digitonin, caused a marked decrease in ecto-ATPase activity; the prior presence of Con A prevented this inhibition, whereas when the lectin was added after the membrane perturbing agent, recovery of the activity was not always possible. The addition of nucleotides before Con A led to a suppression of ecto-ATPase stimulation; it occurred when the nucleotide was hydrolysed (ATP or UTP) and when it was not (adenosine 5′-[β,γ-imido]triphosphate) and even in the presence of 3mM Pi. A model is proposed for the complex regulatory mechanisms of chicken T-tubule ecto-ATPase that involves the occurrence of two different catalytic states in an equilibrium modulated by lectins and cross-linking agents, by the structure of the membrane and by the presence of ligands for a regulatory site.

Expression of avian Ca2+-ATPase in cultured mouse myogenic cells

1989 ◽  
Vol 9 (5) ◽  
pp. 1978-1986
Author(s):  
N J Karin ◽  
Z Kaprielian ◽  
D M Fambrough
Keyword(s):  
Skeletal Muscle ◽  
Endoplasmic Reticulum ◽  
C2c12 Cells ◽  
Cyanine Dye ◽  
Adult Rabbit ◽  
Base Pairs ◽  
Reading Frame ◽  
Cytoplasmic Vesicles ◽  
Fast Twitch ◽  
Chicken Skeletal Muscle

cDNA encoding Ca2+-ATPase was cloned from a chicken skeletal muscle library. The cDNA (termed FCa) comprised 3,239 base pairs, including an open reading frame encoding 994 amino acids which showed the highest degree of homology with the adult rabbit fast-twitch Ca2+-ATPase isoform (C. J. Brandl, S. de Leon, D. R. Martin, and D. H. MacLennan, J. Biol. Chem. 262:3768-3774, 1987). Radiolabeled FCa hybridized to a 3.2-kilobase transcript in chicken skeletal muscle RNA but not to cardiac muscle RNA, which confirmed its identity as encoding the fast Ca2+-ATPase isoenzyme. FCa was transfected into the mouse myogenic line C2C12, from which a protein of 100 kilodaltons was immunopurified by using a monoclonal antibody specific for the avian fast Ca2+-ATPase. Immunofluorescence microscopy of a line (designated C2FCa2) stably expressing the avian Ca2+-ATPase localized the protein to the nuclear envelope and a population of cytoplasmic vesicles. A similar pattern was observed when C2FCa2 cells were stained with DiOC6(3), a cyanine dye that labels endoplasmic reticulum and mitochondria (M. Terasaki, J. Song, J. R. Wong, M. J. Weiss, and L. B. Chen, Cell 38:101-108, 1984). We conclude that the avian Ca2+-ATPase fast isoform is expressed and correctly targeted to the endoplasmic reticulum in mouse C2C12 cells.

The chicken skeletal muscle alpha-actin promoter is tissue specific in transgenic mice

1989 ◽  
Vol 9 (9) ◽  
pp. 3785-3792
Author(s):  
C J Petropoulos ◽  
M P Rosenberg ◽  
N A Jenkins ◽  
N G Copeland ◽  
S H Hughes
Keyword(s):  
Skeletal Muscle ◽  
Transgenic Mice ◽  
Striated Muscle ◽  
Transcriptional Start Site ◽  
Actin Gene ◽  
Tissue Specific ◽  
Adult Mice ◽  
Transgenic Lines ◽  
Alpha Actin ◽  
Chicken Skeletal Muscle

We have generated transgenic mouse lines that carry the promoter region of the chicken skeletal muscle alpha (alpha sk) actin gene linked to the bacterial chloramphenicol acetyltransferase (CAT) gene. In adult mice, the pattern of transgene expression resembled that of the endogenous alpha sk actin gene. In most of the transgenic lines, high levels of CAT activity were detected in striated muscle (skeletal and cardiac) but not in the other tissues tested. In striated muscle, transcription of the transgene was initiated at the normal transcriptional start site of the chicken alpha sk actin gene. The region from nucleotides -191 to +27 of the chicken alpha sk actin gene was sufficient to direct the expression of CAT in striated muscle of transgenic mice. These observations suggest that the mechanism of tissue-specific actin gene expression is well conserved in higher vertebrate species.

scholarly journals Mitochondrial Oxidative Damage in Chicken Skeletal Muscle Induced by Acute Heat Stress

2007 ◽  
Vol 44 (4) ◽  
pp. 439-445 ◽  
Author(s):  
Ahmad Mujahid ◽  
Neil R. Pumford ◽  
Walter Bottje ◽  
Kiyotaka Nakagawa ◽  
Teruo Miyazawa ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Heat Stress ◽  
Oxidative Damage ◽  
Acute Heat Stress ◽  
Mitochondrial Oxidative Damage ◽  
Chicken Skeletal Muscle

scholarly journals Biosynthesis of titin in cultured skeletal muscle cells.

1989 ◽  
Vol 109 (5) ◽  
pp. 2189-2195 ◽  
Author(s):  
W B Isaacs ◽  
I S Kim ◽  
A Struve ◽  
A B Fulton
Keyword(s):  
Skeletal Muscle ◽  
Muscle Cells ◽  
Skeletal Muscle Cells ◽  
Significant Progress ◽  
Large Protein ◽  
Time Period ◽  
And Function ◽  
Triton X ◽  
Developing Muscle ◽  
Synchronized Cultures

Although significant progress has been made regarding the structure and function of titin, little data exist on the biosynthesis of this large protein in developing muscle. Using pulse-labeling with [35S]methionine and immunoprecipitation with an anti-titin mAb, we have examined the biosynthesis of titin in synchronized cultures of skeletal muscle cells derived from day 12 chicken embryos. We find that: (a) titin synthesis increases greater than 4-fold during the first week in culture and during this same time period, synthesis of muscle-specific myosin heavy chain increases greater than 12-fold; (b) newly synthesized titin has a t1/2 of approximately 70 h; (c) titin is resistant to extraction with Triton X-100 both during and immediately after its synthesis. These observations suggest that newly synthesized titin molecules are stable proteins that rapidly associate with the cytoskeleton of developing myotubes.

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