Beef muscle troponin: evidence for multiple forms of troponin-t

Clarke, Frank M.; Lovell, Stephen J.; Masters, C.J.; Winzor, Donald J.

doi:10.1016/0005-2795(76)90205-1

Cited by 34 publications

(21 citation statements)

References 26 publications

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“…The antibody used in this study seems to label two very closely spaced bands corresponding to intact troponin-T. This is likely due to isoforms of troponin-T that are known to exist in skeletal muscle (Briggs et al, 1990;Malhotra, 1994), including specifically bovine skeletal muscle (Clarke et al, 1976; and see discussion in Ho et al, 1994). Both the appearance of the 30-and 28-kDa bands and the disappearance of the intact troponin-T in the MF in this study seem to be very strongly related to the shear force (LSF vs HSF) classification of the original samples.…”

Section: Troponin-tmentioning

confidence: 87%

Proteolysis of specific muscle structural proteins by mu-calpain at low pH and temperature is similar to degradation in postmortem bovine muscle.

Huff-Lonergan

Mitsuhashi

Beekman

et al. 1996

Journal of Animal Science

406

295

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Postmortem (PM) and mu-calpain-induced degradation of specific skeletal muscle proteins was monitored by SDS-PAGE and Western blotting. Samples were removed from bovine longissimus thoracis (LT) at approximately 45 min PM for the preparation of at-death (0-d) myofibrils (MF). The LT was excised at 1 d PM, vacuum-packaged, and stored at 2 degrees C. Samples were removed for Warner-Bratzler shear force analysis and biochemical analysis at 1, 3, 7, 14, 28, and 56 d PM. The protease mu-calpain was purified from bovine skeletal muscle and used to digest at-death MF at pH 5.6, 4 degrees C, 100 microM CaCl2. Degradation of the proteins titin, nebulin, filamin, desmin, and troponin-T was monitored in the PM and mucalpain-digested samples by using SDS-PAGE and Western blotting. The PM samples that had significantly lower shear force (LSF) values (P < .05) at 1 d PM exhibited faster degradation of these five proteins than the higher shear force (HSF) samples. In LSF samples, the intact titin band (T1) was absent by 7 d PM and nebulin was absent by 3 d PM. In LSF samples, some filamin was degraded by 3 d PM, but in HSF samples degradation was not apparent until 14 d PM. In LSF samples, desmin was degraded more rapidly PM than in HSF samples. Troponin-T was broken down PM to yield two major polypeptides of approximately 28 and 30 kDa; these polypeptides appeared earlier PM in LSF samples. Degradation products, similar to those observed PM, for all five proteins also were detected in Western blots of mu-calpain-digested MF, suggesting the calpain system plays a key role in PM protein degradation. ABSTRACT:Postmortem (PM) and m-calpaininduced degradation of specific skeletal muscle proteins was monitored by SDS-PAGE and Western blotting. Samples were removed from bovine longissimus thoracis ( L T ) at approximately 45 min PM for the preparation of at-death (0-d) myofibrils (MF). The LT was excised at 1 d PM, vacuum-packaged, and stored at 2°C. Samples were removed for WarnerBratzler shear force analysis and biochemical analysis at 1, 3, 7, 14, 28, and 56 d PM. The protease m-calpain was purified from bovine skeletal muscle and used to digest at-death MF at pH 5.6, 4°C, 100 mM CaCl 2 . Degradation of the proteins titin, nebulin, filamin, desmin, and troponin-T was monitored in the PM and m-calpain-digested samples by using SDS-PAGE and Western blotting. The PM samples that had significantly lower shear force (LSF) values ( P < .05) at 1 d PM exhibited faster degradation of these five proteins than the higher shear force (HSF) samples. In LSF samples, the intact titin band ( T 1 ) was absent by 7 d PM and nebulin was absent by 3 d PM. In LSF samples, some filamin was degraded by 3 d PM, but in HSF samples degradation was not apparent until 14 d PM. In LSF samples, desmin was degraded more rapidly PM than in HSF samples. Troponin-T was broken down PM to yield two major polypeptides of approximately 28 and 30 kDa; these polypeptides appeared earlier PM in LSF samples. Degradation products, similar to those observed PM, ...

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Section: Troponin-tmentioning

confidence: 87%

Proteolysis of specific muscle structural proteins by mu-calpain at low pH and temperature is similar to degradation in postmortem bovine muscle.

Huff-Lonergan

Mitsuhashi

Beekman

et al. 1996

Journal of Animal Science

406

295

View full text Add to dashboard Cite

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“…Polyacrylamide-gel electrophoresis, in the presence of sodium dodecyl sulfate was performed as previously described (9).…”

Section: Electrophoresismentioning

confidence: 99%

An electron microscope study of the interaction between fructose diphosphate aldolase and actin-containing filaments.

Morton¹,

Clarke²,

Masters³

1977

The Journal of Cell Biology

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The interaction of fructose diphosphate aldolase with F-actin, F-actin-tropomyosin, and F-actin-tropomyosin-troponin has been studied by using negative staining. In the absence of troponin, minor aggregates of aldolase and the F-actin filaments are formed. A well-ordered lattice structure is only formed in the ease of the fully reconstituted filament when the filament-to-filament spacing is 18 nm, and the cross-bridge spacing is 38.7 rim. Evidence is presented that the lattice is due to an interaction between troponin and aldolase. The minimum subunit structure of troponin, still capable of giving rise to a lattice, is the troponin-IT complex, which indicates that troponin-C is not involved in aldolase binding.KEY WORDS actin filaments 9 aldolase tropomyosin 9 troponin

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“…Troponin T (TnT) is an important regulatory and structural component of skeletal muscle thin filaments (Perry, 1998). In bovine muscles, at least two TnT isoforms are present (Clarke et al, 1976). From the fast and slow TnT genes, multiple TnT isoforms are generated by alternative splicing of the messenger RNA in avian and mammalian species (Perry, 1998).…”

Section: Introductionmentioning

confidence: 99%

Amino acid sequences of multiple fast and slow troponin T isoforms expressed in adult bovine skeletal muscles

Muroya

Nakajima

Chikuni

2003

Journal of Animal Science

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Multiple nucleotide sequences of complementary DNA (cDNA) of bovine troponin T (TnT) isoforms expressed in the adult skeletal muscles were determined to facilitate the elucidation of the TnT degradation progress during postmortem aging of muscles. Fresh muscle samples were excised from the lingual, masseter, pectoralis, diaphragm, psoas major, longissimus thoracis, spinnalis, semitendinosus, semimembranosus, and biceps femoris muscles of three Holstein cows within 1 h of slaughter. Complementary DNA fragments of fast and slow TnT isoforms expressed in each muscle were amplified by reverse-transcribed PCR. Consequently, four major fragments of fast TnT and two fragments of slow TnT, all of which contained the complete coding region, were obtained. The sequence determination of these fragments revealed that at least eight and two isoforms were generated by the alternative splicing from bovine fast and slow TnT messenger RNA, respectively. In the fast TnT isoforms, five small variable exons were observed; three of these five exons were in the amino (N)-terminal region. The calculated molecular weight of fast and slow TnT isoforms ranged from 29,816 to 32,125 and from 30,166 to 31,284, respectively. The deduced amino acid sequences revealed that the N-terminal region of all the TnT isoforms was extremely glutamic acid-rich. Reverse-transcribed PCR analysis revealed that expression of each of these isoforms was distributed in a fast or slow muscle-specific manner. Given that TnT degradation has been reported to accompany a decrease in glutamic acid content in the conventional 30-kDa degradation product, the sequence data suggested that the 30-kDa fragment seem to be generated by the proteolytic removal of the glutamic acid-rich N-terminal ends. The multiplicity of TnT isoforms may result in a complicated pattern of TnT degradation on SDS-PAGE gel during beef aging.

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Beef muscle troponin: evidence for multiple forms of troponin-t

Cited by 34 publications

References 26 publications

Proteolysis of specific muscle structural proteins by mu-calpain at low pH and temperature is similar to degradation in postmortem bovine muscle.

Proteolysis of specific muscle structural proteins by mu-calpain at low pH and temperature is similar to degradation in postmortem bovine muscle.

An electron microscope study of the interaction between fructose diphosphate aldolase and actin-containing filaments.

Amino acid sequences of multiple fast and slow troponin T isoforms expressed in adult bovine skeletal muscles

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