Dataset Information

HadBD dehydratase from Mycobacterium tuberculosis FAS-II: a singular structure for a unique function

ABSTRACT: Mycolic acids (MAs) are a unique class of lipids that are essential for viability, virulence and persistence of Mycobacterium tuberculosis (Mtb). Therefore, enzymes involved in MAs biosynthesis represent important class of drug targets. We previously showed that (3R)-hydroxyacyl-ACP dehydratase (HAD) protein HadD is dedicated mainly to the production of keto-MAs and plays a determinant role in Mtb virulence. Here, we discovered that HAD activity requires formation of a tight heterotetramer between HadD and HadB, a HAD unit coded by a distinct chromosomal region. Using biochemical, structural and cell-based analyses, we showed that HadB is the catalytic subunit, whereas HadD is involved in substrate binding. We identified determinants of the ultra-long-chain lipid substrate specificity and revealed details of structure-function relationship. Taken together, our study shows that HadBD, and not HadD, is the biologically relevant functional unit, and these results have important implications for designing innovative antivirulence molecules to fight tuberculosis.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mycobacterium Bovis Bcg Str. Pasteur 1173p2

SUBMITTER: Manuelle Ducoux-Petit

LAB HEAD: Burlet-Schiltz Odile

PROVIDER: PXD043977 | Pride | 2024-03-21

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

Items per page:

1 - 5 of 33

Similar Datasets

Project description:The definition of biomarkers correlating with disease progression could have impact on the rational design of novel diagnostic approaches for bovine tuberculosis in cattle. We have used the advantages of the murine model to identify promising candidates in the host transcriptome post-infection. RNA from BALB/c splenocytes and lung cells after aerogenic Mycobacterium bovis infection were with high-density microarrays to defining biomarkers of disease progression. In antigen-stimulated splenocytes we found statically significant modulation of 1109 genes early after infection and 1134 at later time-point post-infection. 618 of these genes were modulated at both time points. In the lung 282 genes were significantly modulated post-infection. Amongst the most strongly up-regulated genes were granzyme A in spleen, and cxcl9, granzyme B, interleukin-22, interluekin-17A receptor, and ccr6 in lungs. The expression of 20 of the most up-regulated genes identified in the murine studies were evaluated using PBMC from uninfected and naturally infected cattle. We could show that the expression of cxcl9, granzyme A and interleukin-22 following in vitro stimulation with PPD was significantly increased in infected cows compared to naïve animals. Thus, we have demonstrated that murine transcriptome analysis can be used to predict responses in cattle. In addition, we have prioritised CXCL9, GnzA and IL-22 as potential additional readout systems for the ante-mortem diagnosis of bovine tuberculosis. Two groups of 5 BALB/c mice each were infecetd with approx 600 CFU M. bovis via the intranasal route. At days 3 and 14 post challenge five mice per group were euthanized and spleens and lungs harvested. One group of 5 mice were used as a control. Their splenocytes and lung cells were stimulated in vitro for 3 days with M7 protein pool. Antigen cell culture stimulations in mice were performed using an equal pool of seven secreted, immunogenic recombinant mycobacterial proteins (Rv1886c, Rv3019c, Rv3763, Rv3804c, Rv0251, Rv0287 and Rv0288) common to M. bovis and BCG, referred here as M7 protein cocktail. Each protein was used at final concentration of 2μg/ml in 3-day culture.

			Action	DRS
	F098468.dat	Other
	F098470.dat	Other
	F098472.dat	Other
	F098474.dat	Other
	F098476.dat	Other

Dataset Information

HadBD dehydratase from Mycobacterium tuberculosis FAS-II: a singular structure for a unique function

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets