6UW7

The crystal structure of FbiA from Mycobacterium smegmatis, Dehydro-F420-0 bound form

6UW7 の概要

• エントリーDOI: 10.2210/pdb6uw7/pdb
• 関連するPDBエントリー: 6UVX 6UW1 6UW3 6UW5
• 分子名称: Phosphoenolpyruvate transferase, GLYCEROL, 2-[oxidanyl-[(2~{R},3~{S},4~{S})-2,3,4-tris(oxidanyl)-5-[2,4,8-tris(oxidanylidene)-1,9-dihydropyrimido[4,5-b]quinolin-10-yl]pentoxy]phosphoryl]oxyprop-2-enoic acid, ... (5 entities in total)
• 機能のキーワード: factor 420, phosphotransferase, metalloenzyme, biosynthetic protein
• 由来する生物種: Mycolicibacterium smegmatis (strain ATCC 700084 / mc(2)155)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 70358.92

構造登録者

Grinter, R.,Gillett, D.,Cordero, P.R.F.,Izore, T.,Greening, C. (登録日: 2019-11-04, 公開日: 2020-05-13, 最終更新日: 2023-10-11)

主引用文献

Grinter, R.,Ney, B.,Brammananth, R.,Barlow, C.K.,Cordero, P.R.F.,Gillett, D.L.,Izore, T.,Cryle, M.J.,Harold, L.K.,Cook, G.M.,Taiaroa, G.,Williamson, D.A.,Warden, A.C.,Oakeshott, J.G.,Taylor, M.C.,Crellin, P.K.,Jackson, C.J.,Schittenhelm, R.B.,Coppel, R.L.,Greening, C.
Cellular and Structural Basis of Synthesis of the Unique Intermediate Dehydro-F420-0 in Mycobacteria.
mSystems, 5:-, 2020

PubMed Abstract: F is a low-potential redox cofactor used by diverse bacteria and archaea. In mycobacteria, this cofactor has multiple roles, including adaptation to redox stress, cell wall biosynthesis, and activation of the clinical antitubercular prodrugs pretomanid and delamanid. A recent biochemical study proposed a revised biosynthesis pathway for F in mycobacteria; it was suggested that phosphoenolpyruvate served as a metabolic precursor for this pathway, rather than 2-phospholactate as long proposed, but these findings were subsequently challenged. In this work, we combined metabolomic, genetic, and structural analyses to resolve these discrepancies and determine the basis of F biosynthesis in mycobacterial cells. We show that, in whole cells of , phosphoenolpyruvate rather than 2-phospholactate stimulates F biosynthesis. Analysis of F biosynthesis intermediates present in cells harboring genetic deletions at each step of the biosynthetic pathway confirmed that phosphoenolpyruvate is then used to produce the novel precursor compound dehydro-F-0. To determine the structural basis of dehydro-F-0 production, we solved high-resolution crystal structures of the enzyme responsible (FbiA) in apo-, substrate-, and product-bound forms. These data show the essential role of a single divalent cation in coordinating the catalytic precomplex of this enzyme and demonstrate that dehydro-F-0 synthesis occurs through a direct substrate transfer mechanism. Together, these findings resolve the biosynthetic pathway of F in mycobacteria and have significant implications for understanding the emergence of antitubercular prodrug resistance. Mycobacteria are major environmental microorganisms and cause many significant diseases, including tuberculosis. Mycobacteria make an unusual vitamin-like compound, F, and use it to both persist during stress and resist antibiotic treatment. Understanding how mycobacteria make F is important, as this process can be targeted to create new drugs to combat infections like tuberculosis. In this study, we show that mycobacteria make F in a way that is different from other bacteria. We studied the molecular machinery that mycobacteria use to make F, determining the chemical mechanism for this process and identifying a novel chemical intermediate. These findings also have clinical relevance, given that two new prodrugs for tuberculosis treatment are activated by F.

PubMed: 32430409
DOI: 10.1128/mSystems.00389-20

実験手法

X-RAY DIFFRACTION (2.342 Å)