Biosynthesis of muscle‐specific creatine kinase during differentiation in vitro

Morris, Glenn E.; Cole, R. J.

doi:10.1016/0014-5793(77)80379-7

Cited by 7 publications

(1 citation statement)

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“…Perhaps a better indication of the extent of differentiation in culture is the final concentration of M-CK of 0.4-0.6% of the total protein, a value which agrees well with [3Hlleucine-incorporation studies of newly-synthesized M-CK as a proportion of the total newly-synthesized protein in myotube cultures (Morris & Cole, 1977). The value also compares reasonably favourably with the 0.7-1.6% creatine kinase content of 17-19-day embryonic muscle (Table 3).…”

Section: Discussionsupporting

confidence: 66%

Immunoassay of muscle-specific creatine kinase with a monoclonal antibody and application to myogenesis and muscular dystrophy

Morris¹,

Head²

1983

Biochemical Journal

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A competition e.l.i.s.a. (enzyme-linked immunosorbent assay) is described that enables direct measurement of the muscle-specific polypeptide of chick creatine kinase (M-CK) in extracts of differentiating muscle-cell cultures and in blood plasma samples, even in the presence of embryonic, or brain-type, creatine kinase. The characteristics of the assay can be considerably improved by the use of a monoclonal antibody, CK-ART, instead of rabbit antisera, and we offer an explanation for this in terms of heterogeneity of antibody affinities in polyclonal antisera. In addition to native enzyme, the assay will measure creatine kinase unfolded and inactivated by 8 M-urea treatment. During chick muscle differentiation in vitro, M-CK increased from 7.5% of the total creatine kinase at 24h to 76.0% at 143h, in good agreement with isoenzyme separation data. As a percentage of the total cell protein, M-CK increased by 156-340-fold over the same period and constituted 0.38-0.56% of the total protein in late cultures. E.l.i.s.a. measurements on 17-20-day embryonic thigh-muscle extracts, which contain almost exclusively M-CK, agree well with enzyme activity and radioimmunoassay. M-CK constituted 0.7-1.6% of the total protein in 17-19-day embryonic thigh muscle. Plasma M-CK concentrations in normal 2-8-week-old chickens were found to be in the range 0.5-0.9 micrograms/ml. Plasma concentrations of 32-56 micrograms/ml were found in 8-week-old dystrophic chickens by both e.l.i.s.a. and enzyme-activity measurements. The results suggest that inactive or unfolded forms of M-CK do not normally exist, in any significant amounts, in cell and tissue extracts or in freshly prepared samples of plasma.

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Section: Discussionsupporting

confidence: 66%