Solution NMR structure of Alr2454 protein from Nostoc sp. strain PCC 7120, Northeast Structural Genomics Consortium Target NsR264
Northeast Structural Genomics Consortium (NESG)
Solution NMR structure of Alr2454 from Nostoc sp. PCC 7120, the first structural representative of Pfam domain family PF11267.
MTGQELRQLLLDKWGYSYDVQFRRTQGKIFLQVMWKYLEQASFPMNETEYQEHLDSVANYLHALGGAVQVKTFITQTKER PRLGKAVSIPLDLGERASEWIILEHHHHHH
Northeast Structural Genomics Consortium
HELIX DETERMINATION METHOD: AUTHOR DETERMINED
THE PROTEIN IS MONOMERIC BY GEL FILTRATION CHROMATOGRAPHY, STATIC LIGHT SCATTERING AND 15N T1/T2 RELAXATION. THE STRUCTURE WAS DETERMINED USING TRIPLE RESONANCE NMR SPECTROSCOPY. SPECTRA FOR BACKBONE AND SIDE CHAIN ASSIGNMENTS WERE OBTAINED ON A 1.7-MM MICROCRYOPROBE AT 600 MHZ. ALL NOESY DATA WERE ACQUIRED AT 800 MHZ USING A 5-MM CRYOPROBE. BACKBONE ASSIGNMENTS WERE MADE USING PINE, AND THE SIDE CHAIN ASSIGNMENTS WERE COMPLETED MANUALLY. AUTOMATIC NOESY ASSIGNMENTS WERE DETERMINED USING CYANA 3.0. BACKBONE (PHI/PSI) DIHEDRAL ANGLE CONSTRAINTS WERE OBTAINED FROM TALOSplus. COMPLETENESS OF NMR ASSIGNMENTS (EXCLUDING C-TERMINAL HIS6): BACKBONE, 99.4%, SIDE CHAIN, 98.3%, AROMATICS, 96.6%, STEREOSPECIFIC METHYL, 100%, STEREOSPECIFIC SIDE CHAIN NH2: 100.0%. FINAL STRUCTURE QUALITY FACTORS (FOR RESIDUES 1 TO 104, PSVS 1.4), WHERE ORDERED RESIDUES [S(PHI) + S(PSI) > 1.8] COMPRISE: 1-100: (A) RMSD (ORDERED RESIDUES): BB, 0.6, HEAVY ATOM, 0.9. (B) MOLPROBITY RAMACHANDRAN STATISTICS FOR ORDERED RESIDUES: MOST FAVORED, 96.8%, ADDITIONALLY ALLOWED, 3.1%, DISALLOWED, 0.1%. (C) PROCHECK SCORES FOR ORDERED RESIDUES (RAW/Z-): PHI-PSI, -0.15/-0.28, ALL, -0.03/-0.18. (D) MOLPROBITY CLASH SCORE (RAW/Z-): 12.51/-0.62 (E) RPF SCORES FOR GOODNESS OF FIT TO NOESY DATA (RESIDUES 1 TO 104): RECALL, 0.976, PRECISION, 0.934, F-MEASURE, 0.955, DP-SCORE, 0.817. (F) NUMBER OF CLOSE CONTACTS PER 20 MODELS: 2. THE FINAL SIX HISTIDINE RESIDUES IN THE C-TERMINAL AFFINITY TAG WERE NOT INCLUDED IN THE STRUCTURE CALCULATIONS AND HAVE BEEN OMITTED FROM THIS DEPOSITION.
2D 1H-13C HSQC high resolution (L/V methyl stereoassignment)
THE FINAL STRUCTURES ARE BASED ON A TOTAL OF 2478 CONFORMATIONALLY-RESTRICTING NOE-DERIVED DISTANCE CONSTRAINTS AND 162 DIHEDRAL ANGLE CONSTRAINTS; 0 HYDROGEN BOND CONSTRAINTS (25.4 CONSTRAINTS PER RESIDUE, 6.8 LONG RANGE CONSTRAINTS PER RESIDUE, COMPUTED FOR RESIDUES 1 TO 104 BY PSVS 1.4). STRUCTURE DETERMINATION WAS PERFORMED ITERATIVELY USING CYANA 3.0. THE 20 STRUCTURES OUT OF 100 WITH THE LOWEST TARGET FUNCTION WERE FURTHER REFINED BY RESTRAINED MOLECULAR DYNAMICS/ENERGY MINIMIZATION IN EXPLICIT WATER (CNS) WITH PARAM19.
1.01 mM [U-100% 13C; U-100% 15N] NsR264, 20 mM ammonium acetate, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 90% H2O/10% D2O
0.567 mM [U-5% 13C; U-100% 15N] NsR264, 20 mM ammonium acetate, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 90% H2O/10% D2O
Brunger, Adams, Clore, Gros, Nilges and Read
Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
novel fold, Structural Genomics, NORTHEAST STRUCTURAL GENOMICS CONSORTIUM, NESG, PSI-BIOLOGY, Protein Structure Initiative, Unknown Function
MTGQELRQLLLDKWGYSYDVQFRRTQGKIFLQVMWKYLEQASFPMNETEYQEHLDSVANYLHALGGAVQVKTFITQTKER PRLGKAVSIPLDLGERASEWII