Isolation and properties of a basic protein from skeletal-muscle sarcoplasm

Scopes, Robert K.

doi:10.1042/bj0980193

Cited by 19 publications

(9 citation statements)

References 9 publications

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“…This enzyme is relatively inefficient as a carbonic anhydrase (Table 1) but occurs in high concentrations in extracts of skeletal muscle ; yields from the purifications of the chicken and sheep muscle enzymes showed that carbonic anhydrase A accounted for 0.05-0.1 "/, of muscle wet weight. Scopes [47,48] has investigated the relative quantities of soluble protein in aqueous extracts of mammalian muscle and has isolated and examined the molecular properties of a protein (F protein) from pig muscle of unknown function [49]. This protein resembled carbonic anhydrase A in terms of nett surface charge (both are basic proteins), molecular weight (both approximately 34000) and yields following purification (approximately 1 mg /g wet weight muscle), and it is possible that they are the same proteins.…”

Section: Discussionmentioning

confidence: 99%

Purification, Molecular Properties and Ontogeny of Carbonic Anhydrase Isozymes. Evidence for A, B and C Isozymes in Avian and Mammalian Tissues

Holmes

1977

Eur J Biochem

105

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Carbonic anhydrase A from sheep and chicken red skeletal muscles and carbonic anhydrases B and C from chicken intestine and red cells respectively have been isolated in a high state of purity by affinity chromatography. The subunit and native molecular weights of sheep carbonic anhydrase A determined by sodium dodecyl sulphate/polyacrylamide gel electrophoresis and Sephadex gel filtration respectively were both approximately 34000. In contrast, carbonic anhydrases A, B and C from chicken exhibited a molecular weight determined by sodium dodecyl sulphate electrophoresis of 30000. The amino acid composition of sheep muscle carbonic anhydrase A was distinct from the B and C isozymes, particularly in basic amino acid content, which was significantly higher for the A isozyme (38 cJ: 24-291mol). The zinc content of sheep carbonic anhydrase A was found to be 0.81 g atom Zn/mol. The specific activities of chicken carbonic anhydrase isozyme differed significantly according to the ratio A:B:C = 1 :6:47. In addition, these isozymes differed in their affinities for sulphonamide. Chicken carbonic anhydrase A was approximately 100 times less sensitive to acetazolamide inhibition than the chicken C isozyme. Ontogenetic studies in cats showed carbonic anhydrase C to be the only form of the enzyme present in mid-term fetal animals, with carbonic anhydrase A appearing in muscle in the late stages, and carbonic anhydrase B appearing in caecum within 2 days of birth.The evidence is consistent with previous proposals for a third locus encoding carbonic anhydrase in mammalian and avian species. This isozyme (A) is a monomeric, zinc metalloenzyme, which differs from the extensively studied B and C isozymes of carbonic anhydrase in terms of catalytic efficiency, amino acid composition, molecular weight (in sheep), time of ontogenic appearance, sulphonamide inhibition and tissue distribution.

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Section: Discussionmentioning

confidence: 99%

Purification, Molecular Properties and Ontogeny of Carbonic Anhydrase Isozymes. Evidence for A, B and C Isozymes in Avian and Mammalian Tissues

Holmes

1977

Eur J Biochem

105

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“…However, it should be emphasized that this latter worker held muscles at 37°C for 4 hr in order to At 0 and 24 hr, respective/y, from pig number P-2 with a 24 hr gross morphology and color rating of 1.0; (5 and 61 At 0 and 24 hr, respectively, from pig number P-3 with a 24 hr gross morphology and color rating of 1.5. Band identification (according to Scopes, 1966): a) creatine kinase (minor band-fastest migrating), b) phosphoghxomutase, c) creatine kinase (major band-slowest migrating), d) triosephosphate isomerase, el sample slot, f) F-protein.…”

Section: Resultsmentioning

confidence: 99%

A Study of the Sarcoplasmic Proteins of Porcine Muscle by Starch Gel Electrophoresis

Borchert

Powrie

Briskey

1969

Journal of Food Science

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SUMMARY– Extensive investigations were conducted on numerous variables in order to establish the most ideal conditions for the starch gel electrophoresis of the proteins of porcine skeletal muscle sarcoplasm. This technique was used to evaluate the characteristics of the sarcoplasm extracted from (a) muscles at various times post‐mortem, (b) muscles which ultimately remained normal in color and gross morphology as well as muscles which became extremely PSE and (c) muscles which had altered color and gross morphology due to post mortem treatment (i.e., normal color and gross morphology due to surface treatment with L‐N2 and PSE characteristics due to post‐mortem incubation at 37°C for 4 hr).There were no discernible differences between the starch gel electrophoretograms of sarcoplasmic proteins extracted from pre‐ and post‐rigor muscle. Likewise, there were no visually detectable differences between the starch gel electrophoretograms of sarcoplasmic proteins extracted from naturally occurring normal and PSE muscles. Furthermore, the preservation of normal color and gross morphology through L‐N, treatment did not give rise to electrophoretic differences when compared with either naturally occurring normal or PSE muscle.Although sarcoplasmic protein solubility diminished in naturally occurring PSE muscle, this loss in soluble protein did not necessarily manifest itself in the preferential denaturation of a specific sarcoplasmic protein. However when muscles were incubated at 37°C for 4 hr to induce the PSE condition they yielded sarcoplasmic extracts with marked electrophoretic reductions in creatine kinase, phosphoglucomutase, triosephosphate isomerase and F‐protein (Scopes, 1965). Although there were no apparent differences in creatine kinase isozymes between samples of muscle which ultimately remained normal or became PSE, the minor (fastest moving) creatine kinase band, as well as the phosphoglucomutase bands, appeared more labile during incubation at 37°C than the major (slowest moving) creatine kinase band.

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“…A difference near the insertion slot, which can be ascribed to a lower content of the heart and the smooth muscles in aldolase (EC 4.1.2.7) and glyceraldehyde phosphate dehydrogenase (EC 1.2.1.12), is also obvious. On the cathodic side, the band of the diaphragm, absent in cutaneus trunci and heart, which probably corresponds to the basic protein isolated from pig longissimus dorsi by Scopes [1966], does not occur in the smooth muscles.…”

Section: Comparison Of the Electrophoretic Patterns Of Cow Stomach Umentioning

confidence: 99%

“…This method has been applied extensively to fish by Tsuyuki and his collaborators [Tsuyuki et al, 1965;Chen and Tsuyuki, 1970] and to some avian species by Baker [1966]. This has been further elaborated by Scopes [1966Scopes [ ,1968, using pig and rabbit skeletal muscle extracts. Scopes [1966Scopes [ , 1968 has taken advantage of the higher resolution obtained by vertical electrophoresis and of the sharpening of the bands achieved by Poulik's discontinuous system of buffers; he has also described methods for detect ing the main glycolytic enzymes.…”

Section: Introductionmentioning

confidence: 99%

“…This has been further elaborated by Scopes [1966Scopes [ ,1968, using pig and rabbit skeletal muscle extracts. Scopes [1966Scopes [ , 1968 has taken advantage of the higher resolution obtained by vertical electrophoresis and of the sharpening of the bands achieved by Poulik's discontinuous system of buffers; he has also described methods for detect ing the main glycolytic enzymes. The muscle patterns are not only speciesspecific; they make it possible to observe intraspecific variations [Uthe et al, 1966;Tsuyuki et al, 1967;Scopes and Gosselin-Rey, 1968] and hybridization phenomena [Aspinwall and Tsuyuki, 1968;Chen and Tsuyuki, 1970].…”

Section: Introductionmentioning

confidence: 99%

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Comparative Study of the Sarcoplasmic Proteins of Vascular and Some Other Cow Smooth and Striated Muscles by Starch-Gel Electrophoresis

Matagne¹,

Hamoir²

1973

J Vasc Res

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To extend the application of starch-gel electrophoresis to the sarcoplasmic proteins of vertebrate smooth muscle, these extracts must be fractionated by acidification at pH 5 and by ammonium sulphate at 45 % saturation in order to remove the structural proteins and some contaminants which impede the resolution. The extracts of six cow muscles – a white skeletal one, a red skeletal one, the cardiac, stomach, uterus, and carotid muscles – have been analyzed by this method. The patterns have been stained for the major glycolytic enzymes. Enolase, creatine kinase, and lactate dehydrogenase (LDH) make discrimination of the smooth and the striated muscles possible. The smooth muscles contain a much lower amount of glycolytic enzymes and a large number of nonglycolytic bands. Their LDH isoenzymic distribution confirms their high aerobicity. Although many bands are common to the three smooth muscles examined, starch-gel electrophoresis is also able to detect protein differences between the smooth vertebrate muscles of the same species.

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Isolation and properties of a basic protein from skeletal-muscle sarcoplasm

Cited by 19 publications

References 9 publications

Purification, Molecular Properties and Ontogeny of Carbonic Anhydrase Isozymes. Evidence for A, B and C Isozymes in Avian and Mammalian Tissues

Purification, Molecular Properties and Ontogeny of Carbonic Anhydrase Isozymes. Evidence for A, B and C Isozymes in Avian and Mammalian Tissues

A Study of the Sarcoplasmic Proteins of Porcine Muscle by Starch Gel Electrophoresis

Comparative Study of the Sarcoplasmic Proteins of Vascular and Some Other Cow Smooth and Striated Muscles by Starch-Gel Electrophoresis

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