NMR Solution structure of membrane associated protein from Bacillus cereus: Northeast Structural Genomics Consortium Target: BcR97A
Northeast Structural Genomics Consortium (NESG)
Hypothetical Membrane Associated Protein BcR97A
MDLNRMGKDEYYVQITVDGKEVHSKADNGQKYKDYEYKLTGFDKDGKEKELEFTAQKNLRKEAFLRVYHSDKKGVSAWEE VKKDELPAKVKEKLGVKLEHHHHHH
Northeast Structural Genomics Consortium
GFT (4,3)D data was acquired for backbone assignments.
The structure was determined using triple resonance NMR spectroscopy. Automated backbone resonance assignments were made using AutoAssign and pattern picker algorithms developed for automated assignments of GFT NMR data. Side chain assignments were completed manually. Automated NOESY assignments were made using AUTOSTRUCTURE and structure solution was determined using AUTOSTRUCTURE and CYANA-2.1. 176 structures were calculated and 20 best conformers were then refined in a shell of water using CNS. Initial Dihedral constriants were obtained from TALOS. The structure calculations were done including the C-terminal tag LEHHHHHH. Completeness of assignments excluding the 8-residue tag are: Backbone ~98.5%, sidechain ~ 95%, stereospecific methyl assignments 100%. The assignments were validated using the AVS software. Final structure quality factors determined using PSVS software: Ordered residues are defined as: 10-22,32-42,47-58,63-69,76-93. (a) RMSD(ordered residues) all Backbone atoms 0.9A; all heavy atoms 1.4A. (b) Ramachandran statistics for all ordered residues: Most favoured regions:85.9%, Additionally allowed regions 14.0%, Generously allowed region:0.2%. (c) Procheck scores for ordered residues (Raw/Z)phi-psi -0.73/-2.56, All:-0.49/-2.90 (d) MolProbity clash score (Raw/Z): 17.11/-1.41. (e) RPFscores for goodness of fit to NOESY data : Recall:0.963 Precision: 0.805F-measure:0.877 DP-score: 0.848
Zimmerman, Moseley, Kulikowski and Montelione
Brunger, Adams, Clore, Gros, Nilges and Read
NESG, BcR97A, Bacillus cereus, Q812L6 protein, GFT NMR, Structural Genomics, PSI-2, Protein Structure Initiative, Northeast Structural Genomics Consortium, MEMBRANE PROTEIN
DLNRMGKDEYYVQITVDGKEVHSKADNGQKYKDYEYKLTGFDKDGKEKELEFTAQKNLRKEAFLRVYHSDKKGVSAWEEV KKDELPAKVKEKLGVK