Affinity-based capture and identification of protein effectors of the growth regulator ppGpp
Name
nihms-1511130.pdf
Description
Accepted version
Size
2.23 MB
Format
Adobe PDF
Checksum (MD5)
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Author(s)
Wang, Boyuan
Dai, Peng
Ding, David
Del Rosario, Amanda M
Grant, Robert A
Pentelute, Bradley L.
Laub, Michael T
Date Issued
December 2018
Journal
Nature Chemical Biology
Publisher
Springer Science and Business Media LLC
Citation
Wang, Boyuan et al. "Affinity-based capture and identification of protein effectors of the growth regulator ppGpp." Nature Chemical Biology 15, 2 (December 2018): 141–150 © 2018, The Author(s), under exclusive licence to Springer Nature America, Inc.
Version
Author's final manuscript
Abstract
The nucleotide ppGpp is a highly conserved regulatory molecule in bacteria that helps tune growth rate to nutrient availability. Despite decades of study, how ppGpp regulates growth remains poorly understood. Here, we developed and validated a capture-compound mass spectrometry approach that identified >50 putative ppGpp targets in Escherichia coli. These targets control many key cellular processes and include 13 enzymes required for nucleotide synthesis. We demonstrated that ppGpp inhibits the de novo synthesis of all purine nucleotides by directly targeting the enzyme PurF. By solving a structure of PurF bound to ppGpp, we designed a mutation that ablates ppGpp-based regulation, leading to dysregulation of purine-nucleotide synthesis following ppGpp accumulation. Collectively, our results provide new insights into ppGpp-based growth control and a nearly comprehensive set of targets for future exploration. The capture compounds developed should also enable the rapid identification of ppGpp targets in any species, including pathogens. Keywords: bacteria; enzymes; microbiology; target identification; x-ray crystallography
MIT Department
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Chemistry
Massachusetts Institute of Technology. Center for Environmental Health Sciences
Koch Institute for Integrative Cancer Research at MIT
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1038/s41589-018-0183-4
