PMID- 35076225
OWN - NLM
STAT- MEDLINE
DCOM- 20220316
LR  - 20220316
IS  - 1554-8937 (Electronic)
IS  - 1554-8929 (Linking)
VI  - 17
IP  - 2
DP  - 2022 Feb 18
TI  - Discovery of Non-Cysteine-Targeting Covalent Inhibitors by Activity-Based 
      Proteomic Screening with a Cysteine-Reactive Probe.
PG  - 340-347
LID - 10.1021/acschembio.1c00824 [doi]
AB  - Covalent inhibitors of enzymes are increasingly appreciated as pharmaceutical 
      seeds, yet discovering non-cysteine-targeting inhibitors remains challenging. 
      Herein, we report an intriguing experience during our activity-based proteomic 
      screening of 1601 reactive small molecules, in which we monitored the ability of 
      library molecules to compete with a cysteine-reactive iodoacetamide probe. One 
      epoxide molecule, F8, exhibited unexpected enhancement of the probe reactivity 
      for glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a rate-limiting glycolysis 
      enzyme. In-depth mechanistic analysis suggests that F8 forms a covalent adduct 
      with an aspartic acid in the active site to displace NAD(+), a cofactor of the 
      enzyme, with concomitant enhancement of the probe reaction with the catalytic 
      cysteine. The mechanistic underpinning permitted the identification of an 
      optimized aspartate-reactive GAPDH inhibitor. Our findings exemplify that 
      activity-based proteomic screening with a cysteine-reactive probe can be used for 
      discovering covalent inhibitors that react with non-cysteine residues.
FAU - Jung, Yejin
AU  - Jung Y
AD  - Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
AD  - Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
FAU - Noda, Naotaka
AU  - Noda N
AD  - Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
AD  - Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
FAU - Takaya, Junichiro
AU  - Takaya J
AD  - Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
AD  - Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
FAU - Abo, Masahiro
AU  - Abo M
AD  - Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
FAU - Toh, Kohei
AU  - Toh K
AD  - Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
FAU - Tajiri, Ken
AU  - Tajiri K
AD  - Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
FAU - Cui, Changyi
AU  - Cui C
AD  - School of Pharmacy, Fudan University, Shanghai 201203, China.
FAU - Zhou, Lu
AU  - Zhou L
AUID- ORCID: 0000-0002-6807-2647
AD  - School of Pharmacy, Fudan University, Shanghai 201203, China.
FAU - Sato, Shin-Ichi
AU  - Sato SI
AUID- ORCID: 0000-0003-3085-419X
AD  - Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
FAU - Uesugi, Motonari
AU  - Uesugi M
AUID- ORCID: 0000-0002-8515-445X
AD  - Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
AD  - School of Pharmacy, Fudan University, Shanghai 201203, China.
AD  - Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, 
      Uji, Kyoto 611-0011, Japan.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20220125
PL  - United States
TA  - ACS Chem Biol
JT  - ACS chemical biology
JID - 101282906
RN  - EC 1.2.1.- (Glyceraldehyde-3-Phosphate Dehydrogenases)
RN  - K848JZ4886 (Cysteine)
SB  - IM
MH  - Catalysis
MH  - Catalytic Domain
MH  - *Cysteine/chemistry
MH  - Glyceraldehyde-3-Phosphate Dehydrogenases/chemistry/metabolism
MH  - *Proteomics
EDAT- 2022/01/26 06:00
MHDA- 2022/03/17 06:00
CRDT- 2022/01/25 12:18
PHST- 2022/01/26 06:00 [pubmed]
PHST- 2022/03/17 06:00 [medline]
PHST- 2022/01/25 12:18 [entrez]
AID - 10.1021/acschembio.1c00824 [doi]
PST - ppublish
SO  - ACS Chem Biol. 2022 Feb 18;17(2):340-347. doi: 10.1021/acschembio.1c00824. Epub 
      2022 Jan 25.