X-ray structures of Na-GST-1 and Na-GST-2 two glutathione s-transferase from the human hookworm Necator americanus

<p>Abstract</p> <p>Background</p> <p>Human hookworm infection is a major cause of anemia and malnutrition of adults and children in the developing world. As part of on-going efforts to control hookworm infection, The Human Hookworm Vaccine Initiative has identified candidate vaccine antigens from the infective L3 larval stages and adult stages of the parasite. Adult stage antigens include the cytosolic glutathione-S-transferases (GSTs). Nematode GSTs facilitate the inactivation and degradation of a variety of electrophilic substrates (drugs) via the nucleophilic addition of reduced glutathione. Parasite GSTs also play significant roles in multi-drug resistance and the modulation of host-immune defense mechanisms.</p> <p>Results</p> <p>The crystal structures of <it>Na</it>-GST-1 and <it>Na</it>-GST-2, two major GSTs from <it>Necator americanus </it>the main human hookworm parasite, have been solved at the resolution limits of 2.4 Å and 1.9 Å respectively. The structure of <it>Na</it>-GST-1 was refined to R-factor 18.9% (R-free 28.3%) while that of <it>Na</it>-GST-2 was refined to R-factor 17.1% (R-free 21.7%). Glutathione usurped during the fermentation process in bound in the glutathione binding site (G-site) of each monomer of <it>Na</it>-GST-2. <it>Na</it>-GST-1 is uncomplexed and its G-site is abrogated by Gln 50. These first structures of human hookworm parasite GSTs could aid the design of novel hookworm drugs.</p> <p>Conclusion</p> <p>The 3-dimensional structures of <it>Na</it>-GST-1 and <it>Na</it>-GST-2 show two views of human hookworm GSTs. While the GST-complex structure of <it>Na</it>-GST-2 reveals a typical GST G-site that of <it>Na</it>-GST-1 suggests that there is some conformational flexibility required in order to bind the substrate GST. In addition, the overall binding cavities for both are larger, more open, as well as more accessible to diverse ligands than those of GSTs from organisms that have other major detoxifying mechanisms. The results from this study could aid in the design of novel drugs and vaccine antigens.</p

Sequence and structural alignment of Nu class GSTs with a Sigma Class GST (HsGST, human GST or hematopoietic prostagladin D synthase 37) and other parasite GSTs (SjGST, 43, (-GST-1) 42

<p><b>Copyright information:</b></p><p>Taken from "X-ray structures of -GST-1 and -GST-2 two glutathione s-transferase from the human hookworm "</p><p>http://www.biomedcentral.com/1472-6807/7/42</p><p>BMC Structural Biology 2007;7():42-42.</p><p>Published online 26 Jun 2007</p><p>PMCID:PMC1924862.</p><p></p> (a) The alignment reveals that firstly N-terminal alpha beta domain is more conserved than the C-terminal alpha domain. Furthermore, -GST-1 has higher sequence identity with HpolGST than -GST-2 and the lowest similarity is with the HsGST. This figure was generated with ESPript [55, 56]. (b) Structural alignment of monomers of Nu class GSTs (-GST-1, magenta; -GST-2, gold; HpolGST, green) with a sigma class GST (HsGST, cyan)

Nu class GSTs a) -GST-2, c) HpolGST have larger binding cavity than sigma class GST b) HsGST

<p><b>Copyright information:</b></p><p>Taken from "X-ray structures of -GST-1 and -GST-2 two glutathione s-transferase from the human hookworm "</p><p>http://www.biomedcentral.com/1472-6807/7/42</p><p>BMC Structural Biology 2007;7():42-42.</p><p>Published online 26 Jun 2007</p><p>PMCID:PMC1924862.</p><p></p> d) Overlay of the cavities reveals the considerable reduction in the active site size between sigma class (blue) and nu class (cyan). The structure of HpolGST is missing a loop in close proximity to the binding cavity and we modeled it as cartoon from the -GST-2 structure. The glutathione in the G-site is shown as red stick model

Crystallization and preliminary X-ray analysis of Na-SAA-2 from the human hookworm parasite Necator americanus

The purification, crystallization and preliminary X-ray diffraction analysis of a surface-associated antigen from the major human hookworm N. americanus is presented

Spliceosome malfunction causes neurodevelopmental disorders with overlapping features

: Pre-mRNA splicing is a highly coordinated process. While its dysregulation has been linked to neurological deficits, our understanding of the underlying molecular and cellular mechanisms remains limited. We implicated pathogenic variants in U2AF2 and PRPF19, encoding spliceosome subunits in neurodevelopmental disorders (NDDs), by identifying 46 unrelated individuals with 23 de novo U2AF2 missense variants (including seven recurrent variants in 30 individuals) and six individuals with de novo PRPF19 variants. Eight U2AF2 variants dysregulated splicing of a model substrate. Neuritogenesis was reduced in human neurons differentiated from human pluripotent stem cells carrying two U2AF2 hyper-recurrent variants. Neural loss of function of the Drosophila orthologs, U2af50 and Prp19, led to lethality, abnormal mushroom body (MB) patterning, and social deficits, differentially rescued by wild-type and mutant U2AF2 or PRPF19. Transcriptome profiling revealed splicing substrates or effectors (including Rbfox1, a third splicing factor), which rescued MB defects in U2af50 deficient flies. Upon re-analysis of negative clinical exomes followed by data sharing, we further identified six NDD patients carrying RBFOX1 missense variants which, by in vitro testing, showed loss of function. Our study implicates three splicing factors as NDD causative genes and establishes a genetic network with hierarchy underlying human brain development and function