N-tau-methylhistidine turnover in skeletal muscle cells measured by GC-MS

M G Thompson, R M Palmer, A Thom, Karen Elise Garden, Gerald Lobley, Alexander Graham Calder

Research output: Contribution to journalArticle

Abstract

A method that employs gas chromatography-mass spectrometry has been developed to measure N-tau-methylhistidine (3-methylhistidine; 3-MH) synthesis and release from skeletal muscle myotubes in vitro. It shows excellent linearity (0.9999) over the range studied (0-4 nmol), high recovery (92.6%), and low coefficient of variation (1.6%). 3-MH release from myotubes was essentially linear over a 96-h incubation, whereas the loss of 3-MH from cell protein accelerated with increasing time, an effect due, at least in part, to decreasing rates of total protein synthesis. When incubated in either glutamine-free or methionine-free medium for 48 h, 3-MH in cell protein and appearing in the medium were greatly reduced compared with the 48-h controls, suggesting that hypertrophy was greatly reduced. Similar but lesser trends were observed with adenosine 3',5'-cyclic monophosphate. In contrast, 12-O-tetradecanoylphorbol-13-acetate (TPA) appeared to both stimulate 3-MH synthesis and inhibit its release during a 48-h incubation. The development of this method facilitates detailed investigation into the mechanisms through which agents such as TPA regulate myofibrillar protein degradation.

Original languageEnglish
Pages (from-to)C1875-C1879
Number of pages5
JournalAmerican Journal of Physiology: Cell Physiology
Volume270
Issue number6
Publication statusPublished - Jun 1996

Keywords

  • phorbol ester
  • adenosine 3',5'-cyclic monophosphate
  • amino acid deletion
  • myofibrillar protein degradation
  • satellite cells
  • gas chromatography mass spectrometry
  • myofibrillar protein breakdown
  • 3-methylhistidine
  • insulin
  • stimulation
  • metabolism
  • myoblasts
  • invivo
  • acid

Cite this

Thompson, M. G., Palmer, R. M., Thom, A., Garden, K. E., Lobley, G., & Calder, A. G. (1996). N-tau-methylhistidine turnover in skeletal muscle cells measured by GC-MS. American Journal of Physiology: Cell Physiology, 270(6), C1875-C1879.

N-tau-methylhistidine turnover in skeletal muscle cells measured by GC-MS. / Thompson, M G ; Palmer, R M ; Thom, A ; Garden, Karen Elise; Lobley, Gerald; Calder, Alexander Graham.

In: American Journal of Physiology: Cell Physiology, Vol. 270, No. 6, 06.1996, p. C1875-C1879.

Research output: Contribution to journalArticle

Thompson, MG, Palmer, RM, Thom, A, Garden, KE, Lobley, G & Calder, AG 1996, 'N-tau-methylhistidine turnover in skeletal muscle cells measured by GC-MS', American Journal of Physiology: Cell Physiology, vol. 270, no. 6, pp. C1875-C1879.
Thompson, M G ; Palmer, R M ; Thom, A ; Garden, Karen Elise ; Lobley, Gerald ; Calder, Alexander Graham. / N-tau-methylhistidine turnover in skeletal muscle cells measured by GC-MS. In: American Journal of Physiology: Cell Physiology. 1996 ; Vol. 270, No. 6. pp. C1875-C1879.
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