Protein Synthesis in Plant Microsomal System

An active microsomal system from 48-h germinating seeds of Vigna sinensis (L.) Savi has now been developed. It can incorporate amino acids into protein under both in vitro and in vivo conditions, provided dithiothreitol (a protective reagent for SH groups) and phenylthiourea (an inhibitor of phenol oxidase) are present in the huifer system for extraction; and provided the assay mixture contains added dithiothreitol. The system consists of microsomes or ribosomes, tRNA or pH 5 fraction and 20 natural amino acids, ATP and an ATPgenerating system and GTP with requirement for Mg ions. The cell fractions possess aminoacyl-RNA synthctase activity as indicated by the aminoacylliydroxamate formation. Microsomal synthesis is stimulated by exogenous tRNA from Escherichia coli or rat liver and sensitive to various inhibitors such as cycloheximide, chloramphenicoi, fusidic acid. The ribosomal transfer reaction has absolute dependence on the microsomal wash, on the crude enzyme from the same particulate source, and on a synthetic messenger. It is greatly suppressed by fusidic ,acid and by cycloheximide.

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