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

Abstract

MbtH-like proteins (MLPs) are required for soluble expression and/or optimal activity of some adenylation (A) domains of nonribosomal peptide synthetases (NRPSs). Since A domains can interact with noncognate MLP partners, we investigated how the function of an A domain, TioK, involved in the biosynthesis of the bisintercalator thiocoraline, is altered by noncognate MLPs. Measuring TioK activity with 12 different MLPs from a variety of bacterial species using a radiometric assay suggested that the A domain substrate promiscuity could be altered by foreign MLPs. Kinetic studies and bioinformatics analysis indicated that MLPs’ functions are not as simple as previously thought.

Authors:
 [1];  [1];  [2];  [3];  [4]; ORCiD logo [1]
  1. Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lee T. Todd, Jr. Building 789 South Limestone St. Lexington KY 40536-0596 USA
  2. University of Washington, Center for Emerging and Re-emerging Infectious Diseases, 750 Republican St. Seattle WA 98109 USA; University of Washington, Seattle Structural Genomics Center for Infectious Diseases, 307 Westlake Avenue N Seattle WA 98109 USA
  3. University of Washington, Seattle Structural Genomics Center for Infectious Diseases, 307 Westlake Avenue N Seattle WA 98109 USA; Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, P. O. Box 999 Richmond WA 99352 USA; School of Molecular Biosciences, Washington State University, P. O. Box 647520 Pullman WA 99164 USA
  4. Department of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky, Biological Sciences Research Bldg 741 South Limestone St. Lexington KY 40536-0509 USA
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1503572
Report Number(s):
PNNL-SA-131568
Journal ID: ISSN 1439-4227
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Journal Name:
ChemBioChem: a European journal of chemical biology
Additional Journal Information:
Journal Volume: 19; Journal Issue: 20; Journal ID: ISSN 1439-4227
Publisher:
ChemPubSoc Europe
Country of Publication:
United States
Language:
English
Subject:
antibiotics, synthethic biology, non-ribosomal peptide synthetases, SSGCID

Citation Formats

Mori, Shogo, Green, Keith D., Choi, Ryan, Buchko, Garry W., Fried, Michael G., and Garneau-Tsodikova, Sylvie. Using MbtH-Like Proteins to Alter the Substrate Profile of a Nonribosomal Peptide Adenylation Enzyme. United States: N. p., 2018. Web. doi:10.1002/cbic.201800240.
Mori, Shogo, Green, Keith D., Choi, Ryan, Buchko, Garry W., Fried, Michael G., & Garneau-Tsodikova, Sylvie. Using MbtH-Like Proteins to Alter the Substrate Profile of a Nonribosomal Peptide Adenylation Enzyme. United States. doi:10.1002/cbic.201800240.
Mori, Shogo, Green, Keith D., Choi, Ryan, Buchko, Garry W., Fried, Michael G., and Garneau-Tsodikova, Sylvie. Thu . "Using MbtH-Like Proteins to Alter the Substrate Profile of a Nonribosomal Peptide Adenylation Enzyme". United States. doi:10.1002/cbic.201800240.
@article{osti_1503572,
title = {Using MbtH-Like Proteins to Alter the Substrate Profile of a Nonribosomal Peptide Adenylation Enzyme},
author = {Mori, Shogo and Green, Keith D. and Choi, Ryan and Buchko, Garry W. and Fried, Michael G. and Garneau-Tsodikova, Sylvie},
abstractNote = {MbtH-like proteins (MLPs) are required for soluble expression and/or optimal activity of some adenylation (A) domains of nonribosomal peptide synthetases (NRPSs). Since A domains can interact with noncognate MLP partners, we investigated how the function of an A domain, TioK, involved in the biosynthesis of the bisintercalator thiocoraline, is altered by noncognate MLPs. Measuring TioK activity with 12 different MLPs from a variety of bacterial species using a radiometric assay suggested that the A domain substrate promiscuity could be altered by foreign MLPs. Kinetic studies and bioinformatics analysis indicated that MLPs’ functions are not as simple as previously thought.},
doi = {10.1002/cbic.201800240},
journal = {ChemBioChem: a European journal of chemical biology},
issn = {1439-4227},
number = 20,
volume = 19,
place = {United States},
year = {2018},
month = {10}
}

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