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Synthesis of chloramphenicol acetyltransferase in a coupled transcription‐translation in vitro system lacking the chaperones DnaK and DnaJ
Author(s) -
Vysokanov Alexander V
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)01213-x
Subject(s) - chloramphenicol acetyltransferase , ribosome , groel , chloramphenicol , acetyltransferase , protein biosynthesis , biochemistry , biology , translation system , enzyme , ribosomal rna , trimer , cell free system , microbiology and biotechnology , chemistry , escherichia coli , in vitro , acetylation , rna , dimer , gene , promoter , antibiotics , gene expression , organic chemistry
A trimeric enzyme chloramphenicol acetyltransferase (CAT 1 ) has been synthesized in the Zubay system genetically depleted from DnaK and DnaJ. Most of CAT formed in the system fail to assemble into an active trimer. Instead CAT is accumulated in either a GroEL‐bound complex or as an inactive monomer. Addition of purified DnaK and DnaJ to the system prior to the start of protein synthesis leads to the increase of the specific activity of formed CAT. A portion of exogenous DnaK and DnaJ added to the system associate with nascent polypeptide chains in the ribosomes. DnaK also comigrates with 50S‐ribosomal subunits.