Isolation and Identification of Membrane
Proteins Binding to Transfer RNA
LIU Quan-Sheng Jin You-Xin LIU Jian-Hua WANG De-Bao
( State Key Laboratory of Molecular Biology,Shanghai Institute of
Biochemistry,the Chinese Academy of Sciences,Shanghai 200031,China
)
LIU Jian-Hua,JIANG De-Sheng
( Shanghai Research Center of Life Sciences,the Chinese Academy of
Sciences,Shanghai 200031,China )
Abstract Crude extract of total
membrane proteins of yeast was obtained by a method of phase separation in
Triton X-114. The membrane extract was applied to the affinity column on which
yeast total tRNAs were covalently coupled to hydrazinyl Sepharose 4B. By
eluting with increasing concentration gradient of (NH4)2SO4,eluted
proteins were found between concentrations of (NH4)2SO4
0.1 mol/L to 0.3 mol/L. By gel mobility-shift assays,it was also observed that
there were more than two mobility-shift bands on the gel electropherogram after
incubation of sample of 32P-labeled tRNA with the eluent proteins
and the reaction mixtures were analyzed by non-denaturing polyacrylamide gel
electrophoresis. These results confirm that these eluted proteins contain two
major tRNA specific binding proteins.
Key Words Membrane proteins；tRNA；tRNA binding protein
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