Chevelkov, Veniamin; Giller, Karin; Becker, Stefan; Lange, Adam
In this report we present site-specific measurements of amide hydrogen-deuterium exchange rates in a protein in the solid state phase by MAS NMR. Employing perdeuteration, proton detection and a high external magnetic field we could adopt the highly efficient Relax-EXSY protocol previously developed for liquid state NMR. According to this method, we measured the contribution of hydrogen exchange on apparent 15N longitudinal relaxation rates in samples with differing D2O buffer content. Differences in the apparent T1 times allowed us to derive exchange rates for multiple residues in the type III secretion system needle protein.
From- To) 03/01/2016 Final 01/28/2013-12/31/2015 4. TITLE AND SUBTITLE Sa. CONTRACT NUMBER An Energy-Dense AI-NaBH4- PEMFC Based Power Generator for...combination of polymer electrolyte membrane fuel cell ( PEMFC ) with a compact hydrogen generator util izing AI-NaBH4 composite fuel. The conditions...ANSI Std. Z39.18 FLORIDA SOLAR ENERGY CENTER. Crl’nrmg EnPrgy lnrll’pendrnr£’ An Energy-Dense Al-NaBH4- PEMFC Based Power Generator for Unmanned
ANTAGONISMO IN VITRO DE Trichoderma harzianum Rifai SOBRE Fusarium oxysporum Schlecht f. sp passiflorae EN MARACUYÁ (Passiflora edulis Sims var. Flavicarpa DEL MUNICIPIO ZONA BANANERA COLOMBIANA ANTAGONISM IN VITRO OF Trichoderma harzianum Rifai AGAINST Fusarium oxysporum Schlecht f. sp passiflorae IN PASSION FRUIT (Passiflora edulis Sims var. Flavicarpa FROM COLOMBIAN BANANERA ZONE MUNICIPALITY
Reinel José Fernández Barbosa
determining the antagonistic capacity of 6 strains of Trichoderma harzianum against the pathogen. 3 commercial strains were evaluated (TCC-001, TCC-005 and TCC-006 and 3 isolated from soil cultivated with palm oil in the Research Center Caribia of Corpoica (TCN-009, TCN-010, TCN-014. These were evaluated under conditions in vitro using the technique of dual cultivation in Petri plates with Agar Sabouraud. Competition for nutritious and space, micoparasitism and Percentage of Radial Growth Inhibition (PRGI was evaluated, incubating them for 10 days at 28 ºC. A totally randomized design settled down, with 13 treatments and 3 repetitions. All the isolates of T. harzianum exceeded in growth to F. oxysporum with a pathogen growth radiu averages (PGR of 7.42 cm in dual culture. While the pathogen showed average radiu of 1.99 cm. TCN-009 and TCC-006 expressed the best results in competition for nutrients and space to grow 4 times faster than F. oxysporum and to reduce 3 times less the PRGI compared with the control, without significant differences between treatments (P=0.0001; also, produced the largest PRGI to 10 days with values of 64.61 and 65.91% respectively. There was no significant difference when comparing commercial and native isolates, but for the indigenous nature of the isolation, TCN-009 is, an in vitro level, more promising in the search for an antagonist in the control F. oxysporum in similar agroclimatic regions to the Bananera Zone Colombian.
Full Text Available The Type Three Secretion System (T3SS, or injectisome, is a macromolecular infection machinery present in many pathogenic Gram-negative bacteria. It consists of a basal body, anchored in both bacterial membranes, and a hollow needle through which effector proteins are delivered into the target host cell. Two different architectures of the T3SS needle have been previously proposed. First, an atomic model of the Salmonella typhimurium needle was generated from solid-state NMR data. The needle subunit protein, PrgI, comprises a rigid-extended N-terminal segment and a helix-loop-helix motif with the N-terminus located on the outside face of the needle. Second, a model of the Shigella flexneri needle was generated from a high-resolution 7.7-Å cryo-electron microscopy density map. The subunit protein, MxiH, contains an N-terminal α-helix, a loop, another α-helix, a 14-residue-long β-hairpin (Q51-Q64 and a C-terminal α-helix, with the N-terminus facing inward to the lumen of the needle. In the current study, we carried out solid-state NMR measurements of wild-type Shigella flexneri needles polymerized in vitro and identified the following secondary structure elements for MxiH: a rigid-extended N-terminal segment (S2-T11, an α-helix (L12-A38, a loop (E39-P44 and a C-terminal α-helix (Q45-R83. Using immunogold labeling in vitro and in vivo on functional needles, we located the N-terminus of MxiH subunits on the exterior of the assembly, consistent with evolutionary sequence conservation patterns and mutagenesis data. We generated a homology model of Shigella flexneri needles compatible with both experimental data: the MxiH solid-state NMR chemical shifts and the state-of-the-art cryoEM density map. These results corroborate the solid-state NMR structure previously solved for Salmonella typhimurium PrgI needles and establish that Shigella flexneri and Salmonella typhimurium subunit proteins adopt a conserved structure and orientation in their