Activation of an alkaline proteinase from fish skeletal muscle by fatty acids and sodium dodecyl sulphate

A multicatalytic proteinase from rat skeletal muscle contains active site(s) catalysing the degradation of benzoyl-Val-Gly-Arg 4-methyl-7-coumarylamide, succinyl-Ala-Ala-Phe 4-methylcoumarylamide and [14C]methylcasein as well as benzyloxy-carbonyl-Leu-Leu-Glu 2-naphthylamide. These activities are 7-14-fold activated by 1 mM-sodium dodecyl sulphate. The activation leads to a higher susceptibility to the proteinase inhibitor chymostatin and to a lower ability to be inhibited and precipitated by antibodies raised against the non-activated enzyme. Since no changes in Mr or subunit composition were observed in the SDS-activated form, some conformational changes seem to occur during the activation step. More pronounced activation was observed in the presence of physiological concentrations of fatty acids; oleic acid at 100 microM concentrations stimulated the proteinase about 50-fold. In contrast with the non-activated proteinase, the activated enzyme considerably degrades muscle cytoplasmic proteins in vitro. Thus it is not unlikely that, in vivo, potential activators such as fatty acids can induce the multicatalytic proteinase to participate in muscle protein breakdown.

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Myosin light-chain phosphatase

Biochemical Journal ◽

10.1042/bj1570687 ◽

1976 ◽

Vol 157 (3) ◽

pp. 687-697 ◽

Cited By ~ 170

Author(s):

M Morgan ◽

S V Perry ◽

J Ottaway

Keyword(s):

Skeletal Muscle ◽

Light Chain ◽

Sodium Dodecyl Sulphate ◽

Myosin Light Chain Kinase ◽

Adenosine Triphosphatase ◽

Myosin Light Chain ◽

Sodium Dodecyl ◽

Affinity Column ◽

Heavy Meromyosin ◽

Myosin Light Chain Phosphatase

1. A method for the isolation of a new enzyme, myosin light-chain phosphatase, from rabbit white skeletal muscle by using a Sepharose-phosphorylated myosin light-chain affinity column is described. 2. The enzyme migrated as a single component on electrophoresis in sodium dodecyl sulphate/polyacrylamide gel at pH7.0, with apparent mol.wt. 70000. 3. The enzyme was highly specific for the phosphorylated P-light chain of myosin, had pH optima at 6.5 and 8.0 and was not inhibited by NaF. 4. A Ca2+-sensitive ‘ATPase’ (adenosine triphosphatase) system consisting of myosin light-chain kinase, myosin light-chain phosphatase and the P-light chain is described. 5. Evidence is presented for a phosphoryl exchange between Pi, phosphorylated P-light chain and myosin light-chain phosphatase. 6. Heavy meromyosin prepared by chymotryptic digestion can be phosphorylated by myosin light-chain kinase. 7. The ATPase activities of myosin and heavy meromyosin, in the presence and absence of F-actin, were not significantly changed (+/- 10%) by phosphorylation of the P-light chain.

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Analysis of cellular fatty acids and proteins by capillary gas chromatography and sodium dodecyl sulphate polyacrylamide gel electrophoresis to differentiate Mycobacterium avium, Mycobacterium intracellulare and Mycobacterium scrofulaceum (MAIS) complex species

Journal of Chromatography B Biomedical Sciences and Applications ◽

10.1016/s0378-4347(00)83772-x ◽

1990 ◽

Vol 532 ◽

pp. 209-216 ◽

Cited By ~ 4

Author(s):

J. Fourche ◽

M. Capdepuy ◽

J. Maugein ◽

F. Le Moigne

Keyword(s):

Fatty Acids ◽

Gas Chromatography ◽

Mycobacterium Avium ◽

Sodium Dodecyl Sulphate ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Complex Species ◽

Mycobacterium Intracellulare ◽

Cellular Fatty Acids

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Biological activities of bovine cardiac-muscle troponin C C-terminal peptide (residues 84-161)

Biochemical Journal ◽

10.1042/bj2070185 ◽

1982 ◽

Vol 207 (2) ◽

pp. 185-192 ◽

Cited By ~ 5

Author(s):

N V Barskaya ◽

N B Gusev

Keyword(s):

Skeletal Muscle ◽

Amino Acid ◽

Cardiac Muscle ◽

Sodium Dodecyl Sulphate ◽

Binding Sites ◽

Binding Constant ◽

Troponin I ◽

Biological Activities ◽

Sodium Dodecyl ◽

Troponin C

1. Bovine cardiac-muscle troponin C was digested at cysteine residues 35 and 84, and the C-terminal peptide (residues 84-161) was isolated. 2. The C-terminal peptide contains two Ca2+-binding sites. These sites bind Ca2+ with a binding constant of 2.0×10(8) M-1. In the presence of 2 mM-Mg2+ the binding constant for Ca2+ is decreased to 3.70×10(7) M-1. The corresponding constants for native troponin C are 5.90×10(7) M-1. and 2.90×10(7) M-1 respectively. 3. Electrophoretic mobility of the C-terminal peptide is increased in the presence of 0.1 mM-CaCl2 as compared with the mobility in the presence of 2mM-EDTA. The same phenomenon was observed when electrophoresis was performed in the presence of 6 M-urea or 0.1% sodium dodecyl sulphate. 4. When saturated with Ca2+, the C-terminal peptide forms complexes with bovine cardiac-muscle troponin I both in the absence and in the presence of 6 M-urea. This complex is dissociated on removal of Ca2+. 5. The data suggest that the C-terminal peptide of troponin C contains two Ca2+/Mg2+-binding sites and interacts with troponin I. Thus, despite the 30% difference in amino acid composition, the properties of bovine cardiac-muscle troponin C C-terminal peptide are similar to those of rabbit skeletal-muscle troponin C C-terminal peptide.

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The effect of dantrolene on skeletal-muscle sarcoplasmic-reticulum function in malignant hyperpyrexia in pigs

Biochemical Journal ◽

10.1042/bj2120399 ◽

1983 ◽

Vol 212 (2) ◽

pp. 399-405 ◽

Cited By ~ 27

Author(s):

M D White ◽

J G Collins ◽

M A Denborough

Keyword(s):

Skeletal Muscle ◽

Sarcoplasmic Reticulum ◽

Atpase Activity ◽

Muscle Relaxant ◽

Sodium Dodecyl Sulphate ◽

Sodium Dodecyl ◽

Polyacrylamide Gel ◽

Malignant Hyperpyrexia ◽

Dependent Atpase ◽

Sarcoplasmic Reticulum Membrane

The effect of the muscle relaxant dantrolene on isolated sarcoplasmic reticulum was studied in control and malignant-hyperpyrexia-susceptible Landrace pigs. The membranes prepared from both sources showed similar Ca2+-dependent ATPase activities, had comparable phospholipid/protein ratios, and their sodium dodecyl sulphate/polyacrylamide-gel patterns were indistinguishable. Membranes from both sources appeared to bind similar amounts of dantrolene. The drug did not stimulate Ca2+-dependent ATPase activity in preparations from either source. The rates of Ca2+ exchange and Ca2+ efflux appeared to be similar in sarcoplasmic reticulum of control and malignant-hyperpyrexia-susceptible pigs. Dantrolene did not affect either the rates or the amount of Ca2+ lost from the vesicles. These results suggest that dantrolene does not directly affect the movement of Ca2+ across the sarcoplasmic-reticulum membrane.

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A phosphorylated light-chain component of myosin from skeletal muscle

Biochemical Journal ◽

10.1042/bj1350151 ◽

1973 ◽

Vol 135 (1) ◽

pp. 151-164 ◽

Cited By ~ 231

Author(s):

W. T. Perrie ◽

L. B. Smillie ◽

S. V. Perry

Keyword(s):

Skeletal Muscle ◽

Molecular Weight ◽

Amino Acid ◽

Light Chain ◽

Sodium Dodecyl Sulphate ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Low Molecular Weight ◽

Obvious Similarity

1. The low-molecular-weight components of myosin from rabbit skeletal muscle migrated as four bands on polyacrylamide-gel electrophoresis in 8m-urea but only as three in systems containing sodium dodecyl sulphate. The two bands of intermediate mobility in 8m-urea (Ml2 and Ml3) had identical mobilities in sodium dodecyl sulphate. 2. The isolation of pure samples of all four low-molecular-weight components by DEAE-Sephadex chromatography is described. 3. The amino acid compositions of components Ml2 and Ml3 were identical. Further analyses showed the presence of 1 mol of phosphate/18500g of component Ml2 and less than 10% of this amount in component Ml3. Neither light component contained ribose. 4. Alkaline phosphatase from Escherichia coli converted component Ml2 into Ml3. Incubation with crude preparations of phosphorylase b kinase or protein kinase in the presence of ATP converted component Ml3 into Ml2. 5. Phosphorylation of component Ml3 with the kinases isolated from skeletal muscle and [γ-32P]ATP gave incorporation of 32P only into component Ml2 whether whole myosin or separated low-molecular-weight components were used. 6. High-voltage electrophoresis at pH6.5 and pH1.8 of a chymotryptic digest of 32P-labelled component Ml2 yielded one major radioactive peptide containing serine phosphate. 7. The amino acid sequence of this peptide was shown to be: Arg-Ala-Ala-Ala-Glu-Gly-Gly-(Ser,Ser(P))-Asn-Val-Phe. This sequence shows no obvious similarity to the site phosphorylated in the conversion of phosphorylase b into phosphorylase a by phosphorylase b kinase. 8. Evidence suggests that in vivo all the 18500-molecular-weight light chain is in the phosphorylated form. The extent of dephosphorylation that occurred during myosin extraction depended on the conditions employed.

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Photochemistry of aromatic ketones in sodium dodecyl sulphate micelles in the presence of unsaturated fatty acids

Journal of the Serbian Chemical Society ◽

10.2298/jsc0402107m ◽

2004 ◽

Vol 69 (2) ◽

pp. 107-115 ◽

Cited By ~ 7

Author(s):

Dejan Markovic

Keyword(s):

Fatty Acids ◽

Sodium Dodecyl Sulphate ◽

Unsaturated Fatty Acids ◽

Flash Photolysis ◽

Laser Flash Photolysis ◽

Sodium Dodecyl ◽

Self Control ◽

Cage Effect ◽

Radical Pairs ◽

Sds Micelles

Laser-flash photolysis has been employed to characterize the behaviour of the free radicals created in the photochemical reaction of benzophenone (BZP), as well as of its lipoidal derivative, benzophenone-4-heptyl-4?-pentanoic acid (BHPA), with chosen unsaturated fatty acids in sodium dodecyl sulphate micelles. The calculated rate constants were used to study the "cage effect" i.e., the recombination of the created radical-pairs (BZP, BHPA ketyl radical - lipid radical) inside the highly limited space of the SDS micelles. The "cage effect" appears to be the dominant event inside SDS micelles, dependent on the structure of both the reactants-precursors. The fractions of the initially created radical-pairs which escape the "cage effect" and exit into the surrounding aqueous phase do not exceed 16 %. This fact is of enormous importance for the self-control of the pathogenic process of lipid peroxidation.

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Identifikasi Daging Tikus Pada Produk Baso Dengan Metode Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE)

YARSI Medical Journal ◽

10.33476/jky.v26i2.394 ◽

2018 ◽

Vol 26 (2) ◽

pp. 058

Author(s):

Anna P. Roswiem ◽

Triayu Septiani

Keyword(s):

Sodium Dodecyl Sulphate ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Polyacrylamide Gel ◽

Sds Page

Bahan baku untuk membuat baso adalah daging hewan, pada umumnya dari daging sapi, ayam, ikan dan babi. Di beberapa daerah di Indonesia terjadi kasus baso tikus. Tujuan penelitian ini adalah menguji ada tidaknya kandungan daging tikus pada produk baso yang dijual di pasar Cempaka Putih-Kecamatan Kramat Jakarta Pusat dan di pedagang baso atau mie baso di sekitar kampus Universitas YARSI Jakarta. Daging adalah protein salah satu metode untuk mengidentifikasi protein adalah metode Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE). Hasil penelitian menunjukkan bahwa dari 6 sampel baso terindikasi ada 2 sampel baso dengan nomor 1 dan 5 yang dibuat dari campuran daging sapi dan tikus; ada 1 sampel baso dengan nomor 6 yang terbuat dari daging tikus; dan 2 sampel baso dengan nomor 2 dan 3 yang terbuat dari campuran sapi dan babi, dan hanya 1 sampel baso dengan nomor sampel 4 yang benar-benar terbuat dari daging sapi.

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