l-Threonine Transaldolase Activity Is Enabled by a Persistent Catalytic Intermediate.
Amino Acid Sequence
Amino Acids
/ chemistry
Catalysis
Catalytic Domain
Crystallization
Crystallography, X-Ray
Glycine Hydroxymethyltransferase
/ metabolism
Kinetics
Light
Molecular Dynamics Simulation
Protein Conformation
Pyridoxal Phosphate
/ chemistry
Quinones
/ chemistry
Spectrophotometry, Ultraviolet
Threonine
/ chemistry
Journal
ACS chemical biology
ISSN: 1554-8937
Titre abrégé: ACS Chem Biol
Pays: United States
ID NLM: 101282906
Informations de publication
Date de publication:
15 01 2021
15 01 2021
Historique:
pubmed:
19
12
2020
medline:
8
7
2021
entrez:
18
12
2020
Statut:
ppublish
Résumé
l-Threonine transaldolases (lTTAs) are a poorly characterized class of pyridoxal-5'-phosphate (PLP) dependent enzymes responsible for the biosynthesis of diverse β-hydroxy amino acids. Here, we study the catalytic mechanism of ObiH, an lTTA essential for biosynthesis of the β-lactone natural product obafluorin. Heterologously expressed ObiH purifies as a mixture of chemical states including a catalytically inactive form of the PLP cofactor. Photoexcitation of ObiH promotes the conversion of the inactive state of the enzyme to the active form. UV-vis spectroscopic analysis reveals that ObiH catalyzes the retro-aldol cleavage of l-threonine to form a remarkably persistent glycyl quinonoid intermediate, with a half-life of ∼3 h. Protonation of this intermediate is kinetically disfavored, enabling on-cycle reactivity with aldehydes to form β-hydroxy amino acids. We demonstrate the synthetic potential of ObiH via the single step synthesis of (2
Identifiants
pubmed: 33337128
doi: 10.1021/acschembio.0c00753
pmc: PMC8331687
mid: NIHMS1728340
doi:
Substances chimiques
Amino Acids
0
Quinones
0
Threonine
2ZD004190S
Pyridoxal Phosphate
5V5IOJ8338
Glycine Hydroxymethyltransferase
EC 2.1.2.1
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
86-95Subventions
Organisme : NIGMS NIH HHS
ID : DP2 GM137417
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM008349
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM008505
Pays : United States
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