PMID- 34856049
OWN - NLM
STAT- MEDLINE
DCOM- 20220303
LR  - 20220303
IS  - 1439-7633 (Electronic)
IS  - 1439-4227 (Linking)
VI  - 23
IP  - 2
DP  - 2022 Jan 19
TI  - Substrate Enolate Intermediate and Mimic Captured in the Active Site of 
      Streptomyces coelicolor Methylmalonyl-CoA Epimerase.
PG  - e202100487
LID - 10.1002/cbic.202100487 [doi]
AB  - Methylmalonyl-CoA epimerase (MMCE) is proposed to use general acid-base 
      catalysis, but the proposed catalytic glutamic acids are highly asymmetrical in 
      the active site unlike many other racemases. To gain insight into the puzzling 
      relationships between catalytic mechanism, structure, and substrate preference, 
      we solved Streptomyces coelicolor MMCE structures with substrate or 
      2-nitropropionyl-CoA, an intermediate/transition state analogue. Both ligand 
      bound structures have a planar methylmalonate/2-nitropropionyl moiety indicating 
      a deprotonated C2 with >/=4 A distances to either catalytic acid. Both glutamates 
      interact with the carboxylate/nitro group, either directly or through other 
      residues. This suggests the proposed catalytic acids sequentially catalyze proton 
      shifts between C2 and carboxylate of the substrate with an enolate intermediate. 
      In addition, our structures provide a platform to design mutations for expanding 
      substrate scope to support combinatorial biosynthesis.
CI  - (c) 2021 Wiley-VCH GmbH.
FAU - Stunkard, Lee M
AU  - Stunkard LM
AUID- ORCID: 0000-0003-4053-9134
AD  - Department of Biochemistry, Purdue University, 175 S. University St., West 
      Lafayette, IN 47907, USA.
FAU - Benjamin, Aaron B
AU  - Benjamin AB
AUID- ORCID: 0000-0003-4387-2434
AD  - Department of Biochemistry, Purdue University, 175 S. University St., West 
      Lafayette, IN 47907, USA.
FAU - Bower, James B
AU  - Bower JB
AUID- ORCID: 0000-0002-7854-4927
AD  - Department of Biochemistry, Purdue University, 175 S. University St., West 
      Lafayette, IN 47907, USA.
FAU - Huth, Tyler J
AU  - Huth TJ
AUID- ORCID: 0000-0002-5017-6480
AD  - Department of Biochemistry, Purdue University, 175 S. University St., West 
      Lafayette, IN 47907, USA.
FAU - Lohman, Jeremy R
AU  - Lohman JR
AUID- ORCID: 0000-0001-8199-2344
AD  - Department of Biochemistry, Purdue University, 175 S. University St., West 
      Lafayette, IN 47907, USA.
LA  - eng
GR  - DE-AC02-06CH11357/Office of Science/
GR  - 085P1000817/Michigan Economic Development Corporation/
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20211209
PL  - Germany
TA  - Chembiochem
JT  - Chembiochem : a European journal of chemical biology
JID - 100937360
RN  - EC 5.1.- (Racemases and Epimerases)
RN  - EC 5.1.99.1 (methylmalonyl-coenzyme A racemase)
SB  - IM
MH  - Catalysis
MH  - Catalytic Domain
MH  - Humans
MH  - Racemases and Epimerases/*metabolism
MH  - Streptomyces coelicolor/*enzymology
MH  - Substrate Specificity
OTO - NOTNLM
OT  - chirality
OT  - conformational changes
OT  - isomerases
OT  - isosteres
OT  - metalloenzymes
EDAT- 2021/12/03 06:00
MHDA- 2022/03/04 06:00
CRDT- 2021/12/02 17:42
PHST- 2021/11/02 00:00 [revised]
PHST- 2021/09/15 00:00 [received]
PHST- 2021/12/03 06:00 [pubmed]
PHST- 2022/03/04 06:00 [medline]
PHST- 2021/12/02 17:42 [entrez]
AID - 10.1002/cbic.202100487 [doi]
PST - ppublish
SO  - Chembiochem. 2022 Jan 19;23(2):e202100487. doi: 10.1002/cbic.202100487. Epub 2021 
      Dec 9.