Structure of the GcpE-HMBPP complex from Thermus thermophilius

Rekittke, Ingo; Warkentin, Eberhard; Jomaa, Hassan

doi:10.1016/j.bbrc.2015.01.088

Cited by 7 publications

(3 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Structural modeling of Arabidopsis HDS and ligand docking placed MEcPP within the TIM barrel structure of the A-domain and supported a diphosphate linkage between MEcPP and R185, K281, and R342. These 3 residues are strictly conserved across known members of the enzyme family and discerned to be involved in ligand binding of bacterial HDSs (16,27). In addition, the observed binding pattern was consistent with ligand interactions previously reported for HDS (IspG) enzymes from Aquifex aeolicus and Thermus thermophilus (22,24,27).…”

Section: Spatial Positioning Of Mutated Residues By Molecular Modelingsupporting

confidence: 85%

Uncovering the functional residues of Arabidopsis isoprenoid biosynthesis enzyme HDS

Wang

Lei

Xiao

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The methylerythritol phosphate (MEP) pathway is responsible for producing isoprenoids, metabolites with essential functions in the bacterial kingdom and plastid-bearing organisms including plants and Apicomplexa. Additionally, the MEP-pathway intermediate methylerythritol cyclodiphosphate (MEcPP) serves as a plastid-to-nucleus retrograde signal. A suppressor screen of the high MEcPP accumulating mutant plant (ceh1) led to the isolation of 3 revertants (designatedRceh1–3) resulting from independent intragenic substitutions of conserved amino acids in the penultimate MEP-pathway enzyme, hydroxymethylbutenyl diphosphate synthase (HDS). The revertants accumulate varying MEcPP levels, lower than that ofceh1, and exhibit partial or full recovery of MEcPP-mediated phenotypes, including stunted growth and induced expression of stress response genes and the corresponding metabolites. Structural modeling of HDS and ligand docking spatially position the substituted residues at the MEcPP binding pocket and cofactor binding domain of the enzyme. Complementation assays confirm the role of these residues in suppressing theceh1mutant phenotypes, albeit to different degrees. In vitro enzyme assays of wild type and HDS variants exhibit differential activities and reveal an unanticipated mismatch between enzyme kinetics and the in vivo MEcPP levels in the correspondingRcehlines. Additional analyses attribute the mismatch, in part, to the abundance of the first and rate-limiting MEP-pathway enzyme, DXS, and further suggest MEcPP as a rheostat for abundance of the upstream enzyme instrumental in fine-tuning of the pathway flux. Collectively, this study identifies critical residues of a key MEP-pathway enzyme, HDS, valuable for synthetic engineering of isoprenoids, and as potential targets for rational design of antiinfective drugs.

show abstract

Section: Spatial Positioning Of Mutated Residues By Molecular Modelingsupporting

confidence: 85%

Uncovering the functional residues of Arabidopsis isoprenoid biosynthesis enzyme HDS

Wang

Lei

Xiao

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…110 With this significant conformational flexibility, dimeric IspG is thought to exist in a constant equilibrium between its open (holo) and closed (ligand-bound) conformations. 111,114 This property ensures the efficient catalysis described below.…”

Section: Chemical Reviewsmentioning

confidence: 95%

“…It thus enables direct contact between the cluster and the MEcPP binding site, which are contributed from separate monomers . With this significant conformational flexibility, dimeric IspG is thought to exist in a constant equilibrium between its open (holo) and closed (ligand-bound) conformations. , This property ensures the efficient catalysis described below.…”

Section: The Methylerythritol Phosphate Pathwaymentioning

confidence: 99%

The Methylerythritol Phosphate Pathway to Isoprenoids

2016

View full text Add to dashboard Cite

Isoprenoids constitute one of the most diverse classes of natural products. As a compound class, they are essential to basic metabolic processes including cell-wall biosynthesis, post-translational protein modifications, and signaling. In addition, isoprenoid secondary metabolites are highly valuable natural products with a wide range of biotechnological applications. The biosynthesis of their two universal building blocks, isopentenyl diphosphate and dimethylallyl diphosphate, was thought to proceed exclusively by way of mevalonate as a key intermediate until a novel pathway involving methylerithritol phosphate (MEP) was discovered in the early 1990s. In this review, we describe the seven enzymes of the MEP pathway, along with their discoveries, three-dimensional structures, and mechanisms. The latter include examples of remarkable enzyme catalysis including an unusual cytidilation reaction and the use of iron-sulfur cluster cofactors in reductive ring opening and hydroxy-group elimination. Furthermore, isoprenoid biosynthesis shows a characteristic species distribution. A brief overview highlights the MEP pathway's potential as a selective drug target, which is absent in humans but essential to the survival of many important bacterial and apicomplexan pathogens.

show abstract

The Competition Between Chemistry and Biology in Assembling Iron–Sulfur Derivatives: Molecular Structures and Electrochemistry. Part VI. {[Fe 4 S 4 ](S γ Cys ) 3 (nonthiolate ligand)} Proteins

Zanello¹,

Corsini²

2017

Reference Module in Chemistry, Molecular Sciences and Chemical Engineering

View full text Add to dashboard Cite

Structure of the GcpE-HMBPP complex from Thermus thermophilius

Cited by 7 publications

References 34 publications

Uncovering the functional residues of Arabidopsis isoprenoid biosynthesis enzyme HDS

Uncovering the functional residues of Arabidopsis isoprenoid biosynthesis enzyme HDS

The Methylerythritol Phosphate Pathway to Isoprenoids

The Competition Between Chemistry and Biology in Assembling Iron–Sulfur Derivatives: Molecular Structures and Electrochemistry. Part VI. {[Fe 4 S 4 ](S γ Cys ) 3 (nonthiolate ligand)} Proteins

Contact Info

Product

Resources

About