1H, 13C and 15N resonance assignment of truncated SUMO from Trypanosoma brucei

SUMO (small ubiquitin-like modifier) plays important roles in diverse processes by posttranslationally modifying many proteins. Here we report the resonance assignment of the truncated SUMO from Trypanosoma brucei.     

1H, 13C, and 15N assignment of the oxidized and reduced forms of T. brucei glutathione peroxidase-type tryparedoxin peroxidase

The cysteine-homologues of glutathione peroxidases in Trypanosoma brucei catalyze the trypanothione/tryparedoxin-dependent reduction of hydroperoxides. We report the ¹H, ¹³C, and ¹⁵N assignment of the oxidized and reduced form of the enzyme by NMR. Major changes between these two forms were only observed for residues close to the catalytic site.     

Sequence-specific 1H, 13C, and 15N resonance assignment of the autophagy-related protein Atg8

The autophagy-related protein Atg8 is important for the formation of autophagosomes as it mediates membrane fusion. To elucidate the solution structure of Atg8 backbone and side chain chemical shifts of Atg8 were assigned as far as possible.     

1H, 15N and 13C backbone resonance assignment of Rv1567c, an integral membrane protein from Mycobacterium tuberculosis

We report here the backbone assignment of Rv1567c, an integral membrane protein from Mycobacterium tuberculosis. The backbone resonance assignments were determined based on triple-resonance experiments with uniformly [¹³C,¹⁵N]-labeled protein in LMPG detergent micelles.     

1H, 15N, 13C resonance assignment of the acyl carrier protein subunit of the Saccharomyces cerevisiae fatty acid synthase

Acyl carrier proteins participate in the synthesis of fatty acids. Here we report the NMR resonances assignment of the acyl carrier protein domain of the Saccharomyces cerevisiae fatty acid synthase which corresponds to the fragment 138A-302L in the primary structure. The assignment will allow performing NMR studies with the aim to investigate the intrinsic dynamics of this protein, and to study the structural changes upon apo-holo transformation in order to unveil the mechanism of binding of the growing acyl chain.     

1H, 13C, and 15N assignment of the muscular LIM protein MLP/CRP3

The family of CRP proteins comprises three members, which are composed of two LIM domains separated by a long linker of more than 50 residues. We determined the structure of the muscle variant, MLP (CRP3), by nuclear magnetic resonance and show that the two LIM domains are independent of each other.     

1H, 13C, and 15N resonance assignment of the C103S mutant of the N-terminal domain of DsbD from Neisseria meningitidis

We report the nearly complete ¹H, ¹³C, and ¹⁵N resonance assignments of the C103S mutant of the N-terminal domain of DsbD from Neisseria meningitides. Secondary structure determination using CSI method leads to the prediction of nine β-sheet parts.     

1H, 15N and 13C resonance assignment of the pair of Factor-I like modules of the complement protein C7

The carboxy terminus of human complement component C7 comprises two Factor I-like Modules (FIMs) which are essential for formation of the Membrane Attack Complex, the terminal pathway of the innate immune system. C7-FIMs is a 16.9 kDa, recombinant, disulphide-rich, protein encompassing this C-terminal domain. Using conventional triple resonance experiments 93% of the ¹H, ¹⁵N and ¹³C assignment has been achieved, accounting for all assignment apart from a flexible N-terminus cloning artefact and an undefined loop. The chemical shifts have been deposited in the BioMagResBank; Accession No. 15996.     

1H, 13C and 15N resonance assignments for a putative ADF/Cofilin from Trypanosoma brucei

Actin-depolymerizing factor (ADF)/cofilin proteins are a family of actin-binding proteins expressed in almost all eukaryotic cells, and play a significant role in regulating actin-filament dynamics. Here we report the resonance assignments of a putative ADF/cofilin from Trypanosoma brucei for further understanding of the relationship between its structure and function.     

1H, 15N and 13C NMR resonance assignment, secondary structure and global fold of the FMN-binding domain of human cytochrome P450

The FMN-binding domain of human NADPH-cytochrome P450 reductase,corresponding to exons 3-;7, has been expressed at high level in anactive form and labelled with ¹³C and ¹⁵N. Mostof the backbone and aliphatic side-chain ¹H, ¹⁵Nand ¹³C resonances have been assigned using heteronucleardouble- and triple-resonance methods, together with a semiautomaticassignment strategy. The secondary structure as estimated from the chemicalshift index and NOE connectivities consists of six -helices and five-strands. The global fold was deduced from the long-range NOEsunambiguously assigned in a 4D ¹³C-resolved HMQC-NOESY-HMQCspectrum. The fold is of the alternating / type, with the five-strands arranged into a parallel -sheet. The secondarystructure and global fold are very similar to those of the bacterialflavodoxins, but the FMN-binding domain has an extra short helix in place ofa loop, and an extra helix at the N-terminus (leading to the membrane anchordomain in the intact P450 reductase). The experimental constraints werecombined with homology modelling to obtain a structure of the FMN-bindingdomain satisfying the observed NOE constraints. Chemical shift comparisonsshowed that the effects of FMN binding and of FMN reduction are largelylocalised at the binding site.     

1H, 15N, 13C resonance assignment of 9.7 M urea-denatured state of the GTPase effector domain (GED) of dynamin

The GTPase effector domain (GED) of dynamin, a multi-domain protein involved in endocytosis, forms a megadalton-sized self-assembly (even at micromolar concentrations) in native conditions in vitro. While such large assemblies have remained inaccessible to detailed NMR structural characterization, till date, a significant recent achievement has been the elucidation of the GED association pathway starting from a Gdn-HCl denatured monomer. Since, the nature of the denaturant has a strong influence on the conformational preferences in the denatured states, and hence on the association pathways, or even on the final assembly, we report here the NMR resonance assignment of 9.7 M urea-denatured GED from Homo sapiens. This will form the basis for the characterization of the association pathways and the final assembly driven by urea dilution. Jeetender Chugh and Shilpy Sharma have contributed equally.