Using MbtH-Like Proteins to Alter the Substrate Profile of a Nonribosomal Peptide Adenylation Enzyme

Shogo Mori; Keith D. Green; Ryan Choi; Garry W. Buchko; Michael G. Fried; Sylvie Garneau-Tsodikova

doi:10.1002/cbic.201800240

Using MbtH-Like Proteins to Alter the Substrate Profile of a Nonribosomal Peptide Adenylation Enzyme

Shogo Mori, Keith D. Green, Ryan Choi, Garry W. Buchko, Michael G. Fried, Sylvie Garneau-Tsodikova

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

Abstract

MbtH-like proteins (MLPs) are required for soluble expression and/or optimal activity of some adenylation (A) domains of nonribosomal peptide synthetases. Because A domains can interact with noncognate MLP partners, how the function of an A domain, TioK, involved in the biosynthesis of the bisintercalator thiocoraline, is altered by noncognate MLPs has been investigated. Measuring TioK activity with 12 different MLPs from a variety of bacterial species by using a radiometric assay suggested that the A domain substrate promiscuity could be altered by foreign MLPs. Kinetic studies and bioinformatics analysis expanded the complexity of MLP functions and interactions.

Original language	English
Pages (from-to)	2186-2194
Number of pages	9
Journal	ChemBioChem
Volume	19
Issue number	20
DOIs	https://doi.org/10.1002/cbic.201800240
State	Published - Oct 18 2018

Bibliographical note

Funding Information:
This work was supported by a NSF CAREER Award MCB-1149427 (to S.G.-T.); the Department of Pharmaceutical Sciences (to S.G.-T.); the Department of Molecular and Cellular Biochemistry at the University of Kentucky (to M.G.F.); and, in part, with Federal funds awarded to the Seattle Structural Genomics Center for Infectious Disease (SSGCID) from the National Institute of Allergy and Infectious Diseases, National Institutes of Health (NIH), Department of Health and Human Services, under contract no. HHSN272201200025C (to S.G.-T. and G.W.B). S.M. is a recipient of a 2018 long-term visit fellowship from the Yamada Science Foundation, Japan. We thank Dr. Robin Stacy for collaborating on this project. We thank Dr. Wenjing Chen for preliminary cloning and expression of TioK with some MLPs.

Publisher Copyright:
© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

Keywords

bioinformatics
biosynthesis
enzymes
kinetics
protein–protein interactions

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Organic Chemistry

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10.1002/cbic.201800240

Cite this

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abstract = "MbtH-like proteins (MLPs) are required for soluble expression and/or optimal activity of some adenylation (A) domains of nonribosomal peptide synthetases. Because A domains can interact with noncognate MLP partners, how the function of an A domain, TioK, involved in the biosynthesis of the bisintercalator thiocoraline, is altered by noncognate MLPs has been investigated. Measuring TioK activity with 12 different MLPs from a variety of bacterial species by using a radiometric assay suggested that the A domain substrate promiscuity could be altered by foreign MLPs. Kinetic studies and bioinformatics analysis expanded the complexity of MLP functions and interactions.",

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note = "Funding Information: This work was supported by a NSF CAREER Award MCB-1149427 (to S.G.-T.); the Department of Pharmaceutical Sciences (to S.G.-T.); the Department of Molecular and Cellular Biochemistry at the University of Kentucky (to M.G.F.); and, in part, with Federal funds awarded to the Seattle Structural Genomics Center for Infectious Disease (SSGCID) from the National Institute of Allergy and Infectious Diseases, National Institutes of Health (NIH), Department of Health and Human Services, under contract no. HHSN272201200025C (to S.G.-T. and G.W.B). S.M. is a recipient of a 2018 long-term visit fellowship from the Yamada Science Foundation, Japan. We thank Dr. Robin Stacy for collaborating on this project. We thank Dr. Wenjing Chen for preliminary cloning and expression of TioK with some MLPs. Publisher Copyright: {\textcopyright} 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim",

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Using MbtH-Like Proteins to Alter the Substrate Profile of a Nonribosomal Peptide Adenylation Enzyme

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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