Mass: 9605.979 Da / Num. of mol.: 1 / Fragment: NUCLEOTIDE BINDING DOMAIN Source method: isolated from a genetically manipulated source Details: THIS IS A RANDOMLY GENERATED PROTEIN. THE PROTEIN WAS ISOLATED BY CYCLES OF IN VITRO EVOLUTIONARY SELECTION, FROM A RANDOM-SEQUENCE LIBRARY OF 6 X 10**12 MRNA-DISPLAYED PROTEINS. EVOLUTION ...Details: THIS IS A RANDOMLY GENERATED PROTEIN. THE PROTEIN WAS ISOLATED BY CYCLES OF IN VITRO EVOLUTIONARY SELECTION, FROM A RANDOM-SEQUENCE LIBRARY OF 6 X 10**12 MRNA-DISPLAYED PROTEINS. EVOLUTION OF THE PROTEINS PRODUCED WAS DIRECTED BY THE ABILITY TO BIND ATP Source: (gene. exp.) SYNTHETIC CONSTRUCT (others) / Description: SYNTHETIC GENE / Plasmid: MODIFIED PET-24D WITH N-HIS-TEV CLEAVAGE SITE / Production host: ESCHERICHIA COLI BL21(DE3) (bacteria)
Mass: 18.015 Da / Num. of mol.: 99 / Source method: isolated from a natural source / Formula: H2O
Compound details
THE ORIGINAL FUNCTIONAL PROTEIN ISOLATED BY KEEFE AND SZOSTAK (NCBI: AAK50879, GI:13958624) ...THE ORIGINAL FUNCTIONAL PROTEIN ISOLATED BY KEEFE AND SZOSTAK (NCBI: AAK50879, GI:13958624) CONSISTED OF 109 AMINO ACID RESIDUES.A SERIES OF CONSTRUCTS WERE PRODUCED TO IDENTIFY THE FUNCTIONAL CORE OF THE PROTEIN TO AID STRUCTURAL STUDIES. THIS IS A RANDOMLY GENERATED PROTEIN. THE PROTEON WAS ISOLATED BY CYCLES OF IN VITRO EVOLUTIONARY SELECTION FROM A RANDOM-SEQUENCE LIBRARY OF 6 X 10**12 MRNA-DISPLAYED PROTEINS. EVOLUTION OF THE PROTEINS PRODUCED WAS DIRECTED BY THE ABILITY TO BIND ATP.
Sequence details
THE ORIGINAL FUNCTIONAL PROTEIN ISOLATED BY KEEFE AND SZOSTAK (NCBI: AAK50879, GI:13958624) ...THE ORIGINAL FUNCTIONAL PROTEIN ISOLATED BY KEEFE AND SZOSTAK (NCBI: AAK50879, GI:13958624) CONSISTED OF 109 AMINO ACID RESIDUES.A SERIES OF CONSTRUCTS WERE PRODUCED TO IDENTIFY THE FUNCTIONAL CORE OF THE PROTEIN TO AID STRUCTURAL STUDIES. THIS ENTRY CORRESPONDS TO CONSTRUCT REFERRED TO AS ANBP. THE ANBP CONSTRUCT CONSISTS OF RESIDUES 1-78 WHICH WAS EXPRESSED WITH AN N-TERMINAL 6-HIS TAG THAT WAS REMOVED BY CLEAVAGE WITH TEV PROTEASE BEFORE BEING CRYSTALLIZED. THIS GIVES AN INSERTION OF TWO RESIDUES, GA, AT THE N-TER). THIS IS A RANDOMLY GENERATED PROTEIN. THE PROTEON WAS ISOLATED BY CYCLES OF IN VITRO EVOLUTIONARY SELECTION FROM A RANDOM-SEQUENCE LIBRARY OF 6 X 10**12 MRNA-DISPLAYED PROTEINS. EVOLUTION OF THE PROTEINS PRODUCED WAS DIRECTED BY THE ABILITY TO BIND ATP. THERE IS NO UNIPROT ID ASSOCIATED WITH THE SEQUENCE PRESENT IN THIS ENTRY AND THEREFORE, THE DBREF RECORDS INDICATE THAT THE SEQUENCE IS MAPPED TO ITSELF.
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Experimental details
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Experiment
Experiment
Method: X-RAY DIFFRACTION / Number of used crystals: 1
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Sample preparation
Crystal
Density Matthews: 4.4 Å3/Da / Density % sol: 72.1 %
Crystal grow
Method: vapor diffusion, hanging drop / pH: 7.5 Details: USING THE HANGING-DROP VAPOR DIFFUSION TECHIQUE. 20MG/ML OF PROTEIN WAS MIXED IN EQUAL VOLUMES WITH A CRYSTALLIZATION BUFFER OF 0.1M TRIS/HCL PH 8.5, 0.2 M SODIUM CITRATE, 30% PEG 400)
Type: MARRESEARCH / Detector: CCD / Date: Nov 15, 2002 / Details: BENT + TOROIDAL
Radiation
Monochromator: SI111 / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelength
Wavelength: 0.95372 Å / Relative weight: 1
Reflection
Resolution: 1.94→30 Å / Num. obs: 12124 / % possible obs: 99.5 % / Redundancy: 9 % / Rmerge(I) obs: 0.069 / Net I/σ(I): 32
Reflection shell
Resolution: 1.94→2 Å / Redundancy: 7 % / Rmerge(I) obs: 0.477 / Mean I/σ(I) obs: 7.3 / % possible all: 99.3
Reflection
*PLUS
Num. measured all: 108633
Reflection shell
*PLUS
Lowest resolution: 2.01 Å / % possible obs: 99.3 %
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Processing
Software
Name
Version
Classification
REFMAC
5.1.24
refinement
DENZO
datareduction
SCALEPACK
datascaling
SHELXD
phasing
Refinement
Method to determine structure: MAD / Resolution: 1.94→30 Å / Cor.coef. Fo:Fc: 0.943 / Cor.coef. Fo:Fc free: 0.921 / SU B: 1.981 / SU ML: 0.058 / Cross valid method: THROUGHOUT / ESU R: 0.112 / ESU R Free: 0.111 / Stereochemistry target values: MAXIMUM LIKELIHOOD Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. RESIDUES FROM BOTH THE N- AND C-TERMINII WERE NOT VISIBLE IN THE ELECTRON DENSITY. THESE RESIDUES WERE OMITTED FROM THE MODEL. THE REFINED ...Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. RESIDUES FROM BOTH THE N- AND C-TERMINII WERE NOT VISIBLE IN THE ELECTRON DENSITY. THESE RESIDUES WERE OMITTED FROM THE MODEL. THE REFINED MODEL CONTAINS RESIDUES 7- 73. RESIDUES 7-10 OF THE N-TERMINUS ARE NOT IN WELL DEFINED ELECTRON DENSITY. UNACCOUNTED SOLVENT DENSITY IS LOCATED CLOSE TO RESIDUES TYR18 AND TRP35
Rfactor
Num. reflection
% reflection
Selection details
Rfree
0.228
578
4.8 %
RANDOM
Rwork
0.2
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obs
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11530
99.6 %
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Solvent computation
Ion probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: BABINET MODEL PLUS MASK