Structural insights into the catalytic mechanism of a sacrificial sulfur insertase of the N-type ATP pyrophosphatase family, LarE
- Michigan State Univ., East Lansing, MI (United States)
- Michigan State Univ., East Lansing, MI (United States); Univ. catholique de Louvain, Louvain-La-Neuve (Belgium)
The lar operon in Lactobacillus plantarum encodes five Lar proteins (LarA/B/C/D/E) that collaboratively synthesize and incorporate a niacin-derived Ni-containing cofactor into LarA, an Ni-dependent lactate racemase. Previous studies have established that two molecules of LarE catalyze successive thiolation reactions by donating the sulfur atom of their exclusive cysteine residues to the substrate. Here, the catalytic mechanism of this very unusual sulfur-sacrificing reaction remains elusive. In this work, we present the crystal structures of LarE in ligand-free and several ligand-bound forms, demonstrating that LarE is a member of the N-type ATP pyrophosphatase (PPase) family with a conserved N-terminal ATP PPase domain and a unique C-terminal domain harboring the putative catalytic site. Structural analysis, combined with structure-guided mutagenesis, leads us to propose a catalytic mechanism that establishes LarE as a paradigm for sulfur transfer through sacrificing its catalytic cysteine residue.
- Research Organization:
- Argonne National Laboratory (ANL), Argonne, IL (United States)
- Sponsoring Organization:
- National Science Foundation (NSF)
- Grant/Contract Number:
- CHE-1516126
- OSTI ID:
- 1419864
- Journal Information:
- Proceedings of the National Academy of Sciences of the United States of America, Vol. 114, Issue 34; ISSN 0027-8424
- Publisher:
- National Academy of SciencesCopyright Statement
- Country of Publication:
- United States
- Language:
- ENGLISH
Web of Science
Functional Models of the Nickel Pincer Nucleotide Cofactor of Lactate Racemase
|
journal | November 2019 |
Functional Models of the Nickel Pincer Nucleotide Cofactor of Lactate Racemase
|
journal | November 2019 |
Targeting adenylate-forming enzymes with designed sulfonyladenosine inhibitors
|
journal | April 2019 |
Biosynthesis of the nickel-pincer nucleotide cofactor of lactate racemase requires a CTP-dependent cyclometallase
|
journal | June 2018 |
A structural perspective on the PP-loop ATP pyrophosphatase family
|
journal | October 2018 |
Similar Records
Lactate Racemase Nickel-Pincer Cofactor Operates by a Proton-Coupled Hydride Transfer Mechanism
Crystallographic characterization of a tri-Asp metal-binding site at the three-fold symmetry axis of LarE