Unknown

Dataset Information

0

Structural basis of nonnatural amino acid recognition by an engineered aminoacyl-tRNA synthetase for genetic code expansion.


ABSTRACT: The genetic code in a eukaryotic system has been expanded by the engineering of Escherichia coli tyrosyl-tRNA synthetase (TyrRS) with the Y37V and Q195C mutations (37V195C), which specifically recognize 3-iodo-L-tyrosine rather than L-tyrosine. In the present study, we determined the 3-iodo-L-tyrosine- and L-tyrosine-bound structures of the 37V195C mutant of the E. coli TyrRS catalytic domain at 2.0-A resolution. The gamma-methyl group of Val-37 and the sulfur atom of Cys-195 make van der Waals contacts with the iodine atom of 3-iodo-L-tyrosine. The Val-37 and Cys-195 side chains are rigidly fixed by the neighboring residues forming the hydrophobic core of the TyrRS. The major roles of the two mutations are different for the 3-iodo-L-tyrosine-selective recognition in the first step of the aminoacylation reaction (the amino acid activation step): the Y37V mutation eliminates the fatal steric repulsion with the iodine atom, and the Q195C mutation reduces the L-tyrosine misrecognition. The structure of the 37V195C mutant TyrRS complexed with an L-tyrosyladenylate analogue was also solved, indicating that the 3-iodo-L-tyrosine and L-tyrosine side chains are similarly discriminated in the second step (the aminoacyl transfer step). These results demonstrate that the amino acid-binding pocket on the 37V195C mutant is optimized for specific 3-iodo-L-tyrosine recognition.

SUBMITTER: Kobayashi T 

PROVIDER: S-EPMC547826 | biostudies-literature | 2005 Feb

REPOSITORIES: biostudies-literature

altmetric image

Publications

Structural basis of nonnatural amino acid recognition by an engineered aminoacyl-tRNA synthetase for genetic code expansion.

Kobayashi Takatsugu T   Sakamoto Kensaku K   Takimura Tetsuo T   Sekine Ryo R   Kelly Vincent P VP   Kamata Kenji K   Nishimura Susumu S   Yokoyama Shigeyuki S  

Proceedings of the National Academy of Sciences of the United States of America 20050125 5


The genetic code in a eukaryotic system has been expanded by the engineering of Escherichia coli tyrosyl-tRNA synthetase (TyrRS) with the Y37V and Q195C mutations (37V195C), which specifically recognize 3-iodo-L-tyrosine rather than L-tyrosine. In the present study, we determined the 3-iodo-L-tyrosine- and L-tyrosine-bound structures of the 37V195C mutant of the E. coli TyrRS catalytic domain at 2.0-A resolution. The gamma-methyl group of Val-37 and the sulfur atom of Cys-195 make van der Waals  ...[more]

Similar Datasets

| S-EPMC5321558 | biostudies-literature
| S-EPMC7387524 | biostudies-literature
| S-EPMC2396676 | biostudies-literature
| S-EPMC6104698 | biostudies-literature
| S-EPMC10411329 | biostudies-literature
| S-EPMC3401359 | biostudies-literature
| S-EPMC7308279 | biostudies-literature
| S-EPMC4825725 | biostudies-literature
| S-EPMC3053990 | biostudies-literature
| S-EPMC6642615 | biostudies-literature