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Title: Functional interactions between posttranslationally modified amino acids of methyl-coenzyme M reductase in Methanosarcina acetivorans

Abstract

The enzyme methyl-coenzyme M reductase (MCR) plays an important role in mediating global levels of methane by catalyzing a reversible reaction that leads to the production or consumption of this potent greenhouse gas in methanogenic and methanotrophic archaea. In methanogenic archaea, the alpha subunit of MCR (McrA) typically contains four to six posttranslationally modified amino acids near the active site. Recent studies have identified enzymes performing two of these modifications (thioglycine and 5-[S]-methylarginine), yet little is known about the formation and function of the remaining posttranslationally modified residues. Here, we provide in vivo evidence that a dedicated S-adenosylmethionine-dependent methyltransferase encoded by a gene we designated methylcysteine modification (mcmA) is responsible for formation of S-methylcysteine in Methanosarcina acetivorans McrA. Phenotypic analysis of mutants incapable of cysteine methylation suggests that the S-methylcysteine residue might play a role in adaption to mesophilic conditions. To examine the interactions between the S-methylcysteine residue and the previously characterized thioglycine, 5-(S)-methylarginine modifications, we generated M. acetivorans mutants lacking the three known modification genes in all possible combinations. Phenotypic analyses revealed complex, physiologically relevant interactions between the modified residues, which alter the thermal stability of MCR in a combinatorial fashion that is not readily predictable from the phenotypesmore » of single mutants. High-resolution crystal structures of inactive MCR lacking the modified amino acids were indistinguishable from the fully modified enzyme, suggesting that interactions between the posttranslationally modified residues do not exert a major influence on the static structure of the enzyme but rather serve to fine-tune the activity and efficiency of MCR.« less

Authors:
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Publication Date:
Research Org.:
Univ. of Illinois at Urbana-Champaign, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1603218
Alternate Identifier(s):
OSTI ID: 1601490; OSTI ID: 1800037
Grant/Contract Number:  
FG02-02ER15296
Resource Type:
Published Article
Journal Name:
PLoS Biology (Online)
Additional Journal Information:
Journal Name: PLoS Biology (Online) Journal Volume: 18 Journal Issue: 2; Journal ID: ISSN 1545-7885
Publisher:
Public Library of Science (PLoS)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Biochemistry & Molecular Biology; Life Sciences & Biomedicine - Other Topics

Citation Formats

Nayak, Dipti D., Liu, Andi, Agrawal, Neha, Rodriguez-Carerro, Roy, Dong, Shi-Hui, Mitchell, Douglas A., Nair, Satish K., Metcalf, William W., and Sass, ed., Henrik. Functional interactions between posttranslationally modified amino acids of methyl-coenzyme M reductase in Methanosarcina acetivorans. United States: N. p., 2020. Web. doi:10.1371/journal.pbio.3000507.
Nayak, Dipti D., Liu, Andi, Agrawal, Neha, Rodriguez-Carerro, Roy, Dong, Shi-Hui, Mitchell, Douglas A., Nair, Satish K., Metcalf, William W., & Sass, ed., Henrik. Functional interactions between posttranslationally modified amino acids of methyl-coenzyme M reductase in Methanosarcina acetivorans. United States. https://doi.org/10.1371/journal.pbio.3000507
Nayak, Dipti D., Liu, Andi, Agrawal, Neha, Rodriguez-Carerro, Roy, Dong, Shi-Hui, Mitchell, Douglas A., Nair, Satish K., Metcalf, William W., and Sass, ed., Henrik. Mon . "Functional interactions between posttranslationally modified amino acids of methyl-coenzyme M reductase in Methanosarcina acetivorans". United States. https://doi.org/10.1371/journal.pbio.3000507.
@article{osti_1603218,
title = {Functional interactions between posttranslationally modified amino acids of methyl-coenzyme M reductase in Methanosarcina acetivorans},
author = {Nayak, Dipti D. and Liu, Andi and Agrawal, Neha and Rodriguez-Carerro, Roy and Dong, Shi-Hui and Mitchell, Douglas A. and Nair, Satish K. and Metcalf, William W. and Sass, ed., Henrik},
abstractNote = {The enzyme methyl-coenzyme M reductase (MCR) plays an important role in mediating global levels of methane by catalyzing a reversible reaction that leads to the production or consumption of this potent greenhouse gas in methanogenic and methanotrophic archaea. In methanogenic archaea, the alpha subunit of MCR (McrA) typically contains four to six posttranslationally modified amino acids near the active site. Recent studies have identified enzymes performing two of these modifications (thioglycine and 5-[S]-methylarginine), yet little is known about the formation and function of the remaining posttranslationally modified residues. Here, we provide in vivo evidence that a dedicated S-adenosylmethionine-dependent methyltransferase encoded by a gene we designated methylcysteine modification (mcmA) is responsible for formation of S-methylcysteine in Methanosarcina acetivorans McrA. Phenotypic analysis of mutants incapable of cysteine methylation suggests that the S-methylcysteine residue might play a role in adaption to mesophilic conditions. To examine the interactions between the S-methylcysteine residue and the previously characterized thioglycine, 5-(S)-methylarginine modifications, we generated M. acetivorans mutants lacking the three known modification genes in all possible combinations. Phenotypic analyses revealed complex, physiologically relevant interactions between the modified residues, which alter the thermal stability of MCR in a combinatorial fashion that is not readily predictable from the phenotypes of single mutants. High-resolution crystal structures of inactive MCR lacking the modified amino acids were indistinguishable from the fully modified enzyme, suggesting that interactions between the posttranslationally modified residues do not exert a major influence on the static structure of the enzyme but rather serve to fine-tune the activity and efficiency of MCR.},
doi = {10.1371/journal.pbio.3000507},
journal = {PLoS Biology (Online)},
number = 2,
volume = 18,
place = {United States},
year = {2020},
month = {2}
}

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https://doi.org/10.1371/journal.pbio.3000507

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