PMID- 36371819
OWN - NLM
STAT- MEDLINE
DCOM- 20221129
LR  - 20221201
IS  - 1090-2120 (Electronic)
IS  - 0045-2068 (Linking)
VI  - 130
DP  - 2023 Jan
TI  - A simple protein histidine kinase activity assay for high-throughput inhibitor 
      screening.
PG  - 106232
LID - S0045-2068(22)00638-1 [pii]
LID - 10.1016/j.bioorg.2022.106232 [doi]
AB  - Bacterial two-component systems (TCSs), which typically consist of a sensor 
      histidine kinase (HK) and a response regulator (RR), have been investigated as 
      attractive antibacterial drug targets. Unfortunately, current HK activity assays 
      based on the quantification of autophosphorylated HKs are hampered by the 
      instability of the phosphohistidine (pHis) product, rendering them ill-suited for 
      high-throughput screenings. To address this challenge, we developed a simple HK 
      activity assay using readily available reagents, which we have termed AUDECY 
      (AUtophosphorylation-DEphosphorylation CYcle assay). Instead of trying to 
      preserve the fragile pHis, we deliberately decomposed it with a pHis-specific 
      phosphatase to constitute an ATPase-like cycle for convenient colorimetric 
      measurements. This kinetic assay was successfully employed for the kinetic 
      characterization of E. coli EnvZ and for high-throughput inhibitor screening of 
      vancomycin-resistant Enterococcus faecium (VRE) VanS, of which histidine kinase 
      activity was hardly detectable with conventional methods. Through the screening, 
      we identified OSU-03012, a potent VanS HK inhibitor, which sensitized VRE toward 
      vancomycin, highlighting the potential of AUDECY in HK inhibitor discovery.
CI  - Copyright (c) 2022 Elsevier Inc. All rights reserved.
FAU - Lee, Donghee
AU  - Lee D
AD  - Department of Chemistry, Ulsan National Institute of Science and Technology 
      (UNIST), Ulsan 44919, South Korea.
FAU - Lee, Yunmi
AU  - Lee Y
AD  - Discovery Biology Department, Antibacterial Resistance Laboratory, Institut 
      Pasteur Korea, Seongnam-si 13488, South Korea.
FAU - Hye Shin, Son
AU  - Hye Shin S
AD  - Department of Chemistry, Ulsan National Institute of Science and Technology 
      (UNIST), Ulsan 44919, South Korea.
FAU - Min Choi, Su
AU  - Min Choi S
AD  - Department of Chemistry, Ulsan National Institute of Science and Technology 
      (UNIST), Ulsan 44919, South Korea.
FAU - Hyeon Lee, Shin
AU  - Hyeon Lee S
AD  - Department of Chemistry, Ulsan National Institute of Science and Technology 
      (UNIST), Ulsan 44919, South Korea.
FAU - Jeong, Seonghun
AU  - Jeong S
AD  - Department of Chemistry, Ulsan National Institute of Science and Technology 
      (UNIST), Ulsan 44919, South Korea.
FAU - Jang, Soojin
AU  - Jang S
AD  - Discovery Biology Department, Antibacterial Resistance Laboratory, Institut 
      Pasteur Korea, Seongnam-si 13488, South Korea. Electronic address: 
      soojin.jang@ip-korea.org.
FAU - Kee, Jung-Min
AU  - Kee JM
AD  - Department of Chemistry, Ulsan National Institute of Science and Technology 
      (UNIST), Ulsan 44919, South Korea. Electronic address: jmkee@unist.ac.kr.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20221102
PL  - United States
TA  - Bioorg Chem
JT  - Bioorganic chemistry
JID - 1303703
RN  - EC 2.7.13.1 (Histidine Kinase)
RN  - 6Q205EH1VU (Vancomycin)
RN  - EC 2.7.- (Protein Kinases)
RN  - 0 (Transcription Factors)
RN  - 0 (Bacterial Proteins)
SB  - IM
MH  - Histidine Kinase/metabolism
MH  - *Vancomycin/metabolism/pharmacology
MH  - *Escherichia coli/metabolism
MH  - Protein Kinases/metabolism
MH  - High-Throughput Screening Assays
MH  - Transcription Factors/metabolism
MH  - Bacterial Proteins/metabolism
OTO - NOTNLM
OT  - Antibacterial
OT  - ESKAPE pathogens
OT  - High-throughput screening
OT  - Histidine kinase
OT  - Vancomycin resistance
COIS- Declaration of Competing Interest The authors declare that they have no known 
      competing financial interests or personal relationships that could have appeared 
      to influence the work reported in this paper.
EDAT- 2022/11/14 06:00
MHDA- 2022/11/30 06:00
CRDT- 2022/11/13 18:13
PHST- 2022/08/25 00:00 [received]
PHST- 2022/10/15 00:00 [revised]
PHST- 2022/10/26 00:00 [accepted]
PHST- 2022/11/14 06:00 [pubmed]
PHST- 2022/11/30 06:00 [medline]
PHST- 2022/11/13 18:13 [entrez]
AID - S0045-2068(22)00638-1 [pii]
AID - 10.1016/j.bioorg.2022.106232 [doi]
PST - ppublish
SO  - Bioorg Chem. 2023 Jan;130:106232. doi: 10.1016/j.bioorg.2022.106232. Epub 2022 
      Nov 2.