超タンパク質の生合成
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The molecular mechanism in the recognition of L-isoleucine and analog amino acids, by isoleucyl-tRNA synthetase, was elucidated by transferred nuclear Overhauser effect analysis of the conformations of amino acids bound to the enzyme. A variety of nonprotein amino acids, including furanomycin, were found to be incorporated into proteins in <I>in vitro</I> system. Furthermore, L-2-aminohexanoic acid (Ahx) was incorporated into human epidermal growth factor (hEGF), by using <I>Escherichia coli</I> strain harboring plasmid pTA 1522. From the successful production of Ahx-substituted hEGF, a basic strategy was established for preparing proteins substituted with nonprotein amino acid (alloprotein). Induction of the <I>phoA</I> promoter of <I>pho</I> regulon and secretion of the product to the periplasm may depress heat shock-like responses and subsequent hydrolysis of the product by cytoplasmic protease.
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