Characterization of glutamyl-tRNA–dependent dehydratases using nonreactive substrate mimics

Bothwell, Ian R.; Cogan, Dillon P.; Kim, Terry; Reinhardt, Christopher J.; van der Donk, Wilfred A.; Nair, Satish K.

doi:10.1073/pnas.1905240116

Title: Characterization of glutamyl-tRNA–dependent dehydratases using nonreactive substrate mimics

Journal Article · Tue Aug 13 00:00:00 EDT 2019 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1905240116· OSTI ID:1593441

^[1];

^[1]; Kim, Terry ^[1];

^[1];

^[1]; Nair, Satish K. ^[1]

Univ. of Illinois at Urbana-Champaign, IL (United States)

The peptide natural product nisin has been used as a food preservative for 6 decades with minimal development of resistance. Nisin contains the unusual amino acids dehydroalanine and dehydrobutyrine, which are posttranslationally installed by class I lanthipeptide dehydratases (LanBs) on a linear peptide substrate through an unusual glutamyl-tRNA–dependent dehydration of Ser and Thr. To date, little is known about how LanBs catalyze the transfer of glutamate from charged tRNA^Glu to the peptide substrate, or how they carry out the subsequent elimination of the peptide-glutamyl adducts to afford dehydro amino acids. In this work, we describe the synthesis of inert analogs that mimic substrate glutamyl-tRNA^Glu and the glutamylated peptide intermediate, and determine the crystal structures of 2 LanBs in complex with each of these compounds. Mutational studies were used to characterize the function of the glutamylation and glutamate elimination active-site residues identified through the structural analysis. These combined studies provide insights into the mechanisms of substrate recognition, glutamylation, and glutamate elimination by LanBs to effect a net dehydration reaction of Ser and Thr.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: National Institutes of Health (NIH)

Grant/Contract Number:: GM058822; GM079038; F32 GM117765

OSTI ID:: 1593441

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, Issue 35; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 30 works

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