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Genetic incorporation of histidine derivatives using an engineered pyrrolysyl-tRNA synthetase.

ABSTRACT: A polyspecific amber suppressor aminoacyl-tRNA synthetase/tRNA pair was evolved that genetically encodes a series of histidine analogues in both Escherichia coli and mammalian cells. In combination with tRNACUA(Pyl), a pyrrolysyl-tRNA synthetase mutant was able to site-specifically incorporate 3-methyl-histidine, 3-pyridyl-alanine, 2-furyl-alanine, and 3-(2-thienyl)-alanine into proteins in response to an amber codon. Substitution of His66 in the blue fluorescent protein (BFP) with these histidine analogues created mutant proteins with distinct spectral properties. This work further expands the structural and chemical diversity of unnatural amino acids (UAAs) that can be genetically encoded in prokaryotic and eukaryotic organisms and affords new probes of protein structure and function.

SUBMITTER: Xiao H

PROVIDER: S-EPMC4033645 | biostudies-literature | 2014 May

REPOSITORIES: biostudies-literature

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Genetic incorporation of histidine derivatives using an engineered pyrrolysyl-tRNA synthetase.

Xiao Han H Peters Francis B FB Yang Peng-Yu PY Reed Sean S Chittuluru Johnathan R JR Schultz Peter G PG

ACS chemical biology 20140317 5

A polyspecific amber suppressor aminoacyl-tRNA synthetase/tRNA pair was evolved that genetically encodes a series of histidine analogues in both Escherichia coli and mammalian cells. In combination with tRNACUA(Pyl), a pyrrolysyl-tRNA synthetase mutant was able to site-specifically incorporate 3-methyl-histidine, 3-pyridyl-alanine, 2-furyl-alanine, and 3-(2-thienyl)-alanine into proteins in response to an amber codon. Substitution of His66 in the blue fluorescent protein (BFP) with these histidi ...[more]

PMID: 24506189

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