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JBC, Vol. 252, Issue 10, 3422-3429, May, 1977
A. F. Martin, M. Rabinowitz, R. Blough, G. Prior and R. Zak
The kinetics of labeling of myosin heavy chain, following a single
intravenous injection of L-[4,5-3H]leucine, were analyzed with the help of
a computer, in conjunction with the labeling kinetics of the specific
radioactivities of the precursor amino acid pool. As precursor we used
leucyl-tRNA which, as we show here, differs significantly from the
intracellular free leucine pool. The half-life of myosin heavy chain was
determined from the initial period (0 to 60 min) of incorporation of label
into protein after a single injection of tritiated leucine, and also from
the period (7 to 14 days) when there is exponential decay of the labeled
protein. Myosin heavy chain was separated from other myofibrillar proteins
by polyacrylamide gel electrophoresis before measurement of leucine
specific radioactivity. The specific radioactivity was measured in both
protein and precursor pools by a sensitive isotope dilution procedure
(range, 100 to 1500 pmol). The values for the half-life of myosin heavy
chain determined at both intervals were similar (5.4 and 5.9 days).
Substitution of the specific radioactivity of the intracellular free
leucine pool decreased the half-life to 2.7 dyas. Similar values were
obtained when the half-life was calculated by simple graphical integration
of the experimental curves.
Measurements of half-life of rat cardiac myosin heavy chain with leucyl-tRNA used as precursor pool
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