HEADER COMPLEX (CALCIUM-BINDING/TRANSFERASE) 23-SEP-97 1CM1 TITLE MOTIONS OF CALMODULIN - SINGLE-CONFORMER REFINEMENT COMPND MOL_ID: 1; COMPND 2 MOLECULE: CALMODULIN; COMPND 3 CHAIN: A; COMPND 4 MOL_ID: 2; COMPND 5 MOLECULE: CALMODULIN-DEPENDENT PROTEIN KINASE II-ALPHA; COMPND 6 CHAIN: B; COMPND 7 FRAGMENT: CALMODULIN BINDING DOMAIN, RESIDUES 290 - 314; COMPND 8 EC: 2.7.1.123; COMPND 9 ENGINEERED: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: BOS TAURUS; SOURCE 3 ORGANISM_COMMON: BOVINE; SOURCE 4 ORGAN: BRAIN; SOURCE 5 OTHER_DETAILS: SIGMA LOT 54H9558; SOURCE 6 MOL_ID: 2; SOURCE 7 SYNTHETIC: YES; SOURCE 8 ORGANISM_SCIENTIFIC: BOS TAURUS; SOURCE 9 ORGANISM_COMMON: BOVINE KEYWDS COMPLEX (CALCIUM-BINDING/TRANSFERASE), EF-HAND KEYWDS 2 CALCIUM-BINDING PROTEIN EXPDTA X-RAY DIFFRACTION AUTHOR M.E.WALL,G.N.PHILLIPS JUNIOR REVDAT 1 04-MAR-98 1CM1 0 JRNL AUTH M.E.WALL,J.B.CLARAGE,G.N.PHILLIPS JUNIOR JRNL TITL MOTIONS OF CALMODULIN CHARACTERIZED USING BOTH JRNL TITL 2 BRAGG AND DIFFUSE X-RAY SCATTERING JRNL REF STRUCTURE (LONDON) V. 5 1599 1997 JRNL REFN ASTM STRUE6 UK ISSN 0969-2126 2005 REMARK 1 REMARK 1 REFERENCE 1 REMARK 1 AUTH W.E.MEADOR,A.R.MEANS,F.A.QUIOCHO REMARK 1 TITL MODULATION OF CALMODULIN PLASTICITY IN MOLECULAR REMARK 1 TITL 2 RECOGNITION ON THE BASIS OF X-RAY STRUCTURES REMARK 1 REF SCIENCE V. 262 1718 1993 REMARK 1 REFN ASTM SCIEAS US ISSN 0036-8075 0038 REMARK 1 REFERENCE 2 REMARK 1 AUTH W.E.MEADOR,A.R.MEANS,F.A.QUIOCHO REMARK 1 TITL TARGET ENZYME RECOGNITION BY CALMODULIN: 2.4 A REMARK 1 TITL 2 STRUCTURE OF A CALMODULIN-PEPTIDE COMPLEX REMARK 1 REF SCIENCE V. 257 1251 1992 REMARK 1 REFN ASTM SCIEAS US ISSN 0036-8075 0038 REMARK 2 REMARK 2 RESOLUTION. 2.0 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : X-PLOR 3.851 REMARK 3 AUTHORS : BRUNGER REMARK 3 REMARK 3 DATA USED IN REFINEMENT. REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 2.0 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 10.0 REMARK 3 DATA CUTOFF (SIGMA(F)) : 2.0 REMARK 3 DATA CUTOFF HIGH (ABS(F)) : 100000. REMARK 3 DATA CUTOFF LOW (ABS(F)) : 0.1 REMARK 3 COMPLETENESS (WORKING+TEST) (%) : 92.1 REMARK 3 NUMBER OF REFLECTIONS : 11522 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 CROSS-VALIDATION METHOD : THROUGHOUT REMARK 3 FREE R VALUE TEST SET SELECTION : RANDOM REMARK 3 R VALUE (WORKING SET) : 0.234 REMARK 3 FREE R VALUE : 0.302 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 10.3 REMARK 3 FREE R VALUE TEST SET COUNT : 1189 REMARK 3 ESTIMATED ERROR OF FREE R VALUE : 0.009 REMARK 3 REMARK 3 FIT IN THE HIGHEST RESOLUTION BIN. REMARK 3 TOTAL NUMBER OF BINS USED : 10 REMARK 3 BIN RESOLUTION RANGE HIGH (A) : 2.0 REMARK 3 BIN RESOLUTION RANGE LOW (A) : 2.07 REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 82.8 REMARK 3 REFLECTIONS IN BIN (WORKING SET) : 964 REMARK 3 BIN R VALUE (WORKING SET) : 0.344 REMARK 3 BIN FREE R VALUE : 0.369 REMARK 3 BIN FREE R VALUE TEST SET SIZE (%) : 11.7 REMARK 3 BIN FREE R VALUE TEST SET COUNT : 128 REMARK 3 ESTIMATED ERROR OF BIN FREE R VALUE : 0.033 REMARK 3 REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT. REMARK 3 PROTEIN ATOMS : 1258 REMARK 3 NUCLEIC ACID ATOMS : 0 REMARK 3 HETEROGEN ATOMS : 5 REMARK 3 SOLVENT ATOMS : 58 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : 22.5 REMARK 3 MEAN B VALUE (OVERALL, A**2) : 41.4 REMARK 3 OVERALL ANISOTROPIC B VALUE. REMARK 3 B11 (A**2) : NULL REMARK 3 B22 (A**2) : NULL REMARK 3 B33 (A**2) : NULL REMARK 3 B12 (A**2) : NULL REMARK 3 B13 (A**2) : NULL REMARK 3 B23 (A**2) : NULL REMARK 3 REMARK 3 ESTIMATED COORDINATE ERROR. REMARK 3 ESD FROM LUZZATI PLOT (A) : 0.29 REMARK 3 ESD FROM SIGMAA (A) : 0.31 REMARK 3 LOW RESOLUTION CUTOFF (A) : 5.00 REMARK 3 REMARK 3 CROSS-VALIDATED ESTIMATED COORDINATE ERROR. REMARK 3 ESD FROM C-V LUZZATI PLOT (A) : 0.34 REMARK 3 ESD FROM C-V SIGMAA (A) : 0.29 REMARK 3 REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES. REMARK 3 BOND LENGTHS (A) : 0.009 REMARK 3 BOND ANGLES (DEGREES) : 2.3 REMARK 3 DIHEDRAL ANGLES (DEGREES) : 21.8 REMARK 3 IMPROPER ANGLES (DEGREES) : 0.72 REMARK 3 REMARK 3 ISOTROPIC THERMAL MODEL : RESTRAINED REMARK 3 REMARK 3 ISOTROPIC THERMAL FACTOR RESTRAINTS. RMS SIGMA REMARK 3 MAIN-CHAIN BOND (A**2) : 2.22 ; 1.50 REMARK 3 MAIN-CHAIN ANGLE (A**2) : 3.88 ; 2.00 REMARK 3 SIDE-CHAIN BOND (A**2) : 3.31 ; 2.00 REMARK 3 SIDE-CHAIN ANGLE (A**2) : 5.47 ; 2.50 REMARK 3 REMARK 3 NCS MODEL : NULL REMARK 3 REMARK 3 NCS RESTRAINTS. RMS SIGMA/WEIGHT REMARK 3 GROUP 1 POSITIONAL (A) : NULL ; NULL REMARK 3 GROUP 1 B-FACTOR (A**2) : NULL ; NULL REMARK 3 REMARK 3 PARAMETER FILE 1 : PARAM19X.PRO REMARK 3 PARAMETER FILE 2 : PARAM19.SOL REMARK 3 TOPOLOGY FILE 1 : TOPH19X.PRO REMARK 3 TOPOLOGY FILE 2 : TOPH19.SOL REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: NULL REMARK 4 REMARK 4 1CM1 COMPLIES WITH FORMAT V. 2.2, 16-DEC-1996 REMARK 6 REMARK 6 THE FIRST THREE RESIDUES OF CALMODULIN (ALA 1, ASP 2, AND REMARK 6 GLN 3) ARE NOT PRESENT IN THE DENSITY AND ARE NOT INCLUDED. REMARK 6 THIS IS ALSO TRUE FOR THE C-TERMINAL RESIDUES ALA 147 AND REMARK 6 LYS 148. THE FIRST THREE AND THE LAST FOUR RESIDUES OF THE REMARK 6 TWENTY-FIVE RESIDUE PEPTIDE ARE ALSO DISORDERED AND ARE NOT REMARK 6 INCLUDED. REMARK 7 REMARK 7 RESIDUES 74 - 83, WHICH ARE NOT PRESENT IN PDB ENTRY 1CDM, REMARK 7 WERE ADDED FOR THIS REFINEMENT. AN INITIAL GUESS WAS REMARK 7 OBTAINED FROM THE COORDINATES OF PDB ENTRY 1CDL. THESE REMARK 7 RESIDUES DO NOT SHOW CONNECTED ELECTRON DENSITY AT A LEVEL REMARK 7 OF 1SIGMA. REMARK 8 REMARK 8 THIS SINGLE-CONFORMER MODEL (1CM1) IS TO BE COMPARED WITH REMARK 8 THE FOUR-CONFORMER MODEL (1CM4). THE DATA DEPOSITED WITH REMARK 8 THIS MODEL WERE ALSO USED TO REFINE 1CM4. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : MAY-1996 REMARK 200 TEMPERATURE (KELVIN) : 302 REMARK 200 PH : 5.2 REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : Y REMARK 200 RADIATION SOURCE : CHESS REMARK 200 BEAMLINE : F2 REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 0.98 REMARK 200 MONOCHROMATOR : SINGLE-CRYSTAL REMARK 200 OPTICS : MIRRORS REMARK 200 REMARK 200 DETECTOR TYPE : CCD REMARK 200 DETECTOR MANUFACTURER : PRINCETON UNIVERSITY REMARK 200 INTENSITY-INTEGRATION SOFTWARE : DENZO REMARK 200 DATA SCALING SOFTWARE : SCALEPACK REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 11522 REMARK 200 RESOLUTION RANGE HIGH (A) : 2.0 REMARK 200 RESOLUTION RANGE LOW (A) : 10.0 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : 2. REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 92.1 REMARK 200 DATA REDUNDANCY : NULL REMARK 200 R MERGE (I) : NULL REMARK 200 R SYM (I) : 0.061 REMARK 200 <I/SIGMA(I)> FOR THE DATA SET : 9.2 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 2.00 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 2.07 REMARK 200 COMPLETENESS FOR SHELL (%) : 82.8 REMARK 200 DATA REDUNDANCY IN SHELL : NULL REMARK 200 R MERGE FOR SHELL (I) : NULL REMARK 200 R SYM FOR SHELL (I) : 0.24 REMARK 200 <I/SIGMA(I)> FOR SHELL : NULL REMARK 200 REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: MOLECULAR REMARK 200 REPLACEMENT REMARK 200 SOFTWARE USED: X-PLOR REMARK 200 STARTING MODEL: PDB ENTRIES 1CDM AND 1CDL REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): NULL REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): NULL REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: DIFFRACTION-QUALITY CRYSTALS REMARK 280 WERE MICROSEEDED IN HANGING DROPS OVER 100 MM SODIUM REMARK 280 ACETATE AT PH 5.2, WITH 20% POLY-ETHYLENE GLYCOL 6000 REMARK 280 (PEG 6000), 10 MM CALCIUM CHLORIDE AND 0.02% SODIUM REMARK 280 AZIDE. STOCK SOLUTIONS OF 24 MG/ML BOVINE BRAIN REMARK 280 CALMODULIN (SIGMA LOT 54H9558), 14 MG/ML CAMKII-ALPHA REMARK 280 PEPTIDE, AND 30% PEG WERE MIXED INTO HANGING DROPS IN REMARK 280 ABOUT A 4-2-1 RATIO. REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: C 2 2 21 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 -X,-Y,1/2+Z REMARK 290 3555 -X,Y,1/2-Z REMARK 290 4555 X,-Y,-Z REMARK 290 5555 1/2+X,1/2+Y,Z REMARK 290 6555 1/2-X,1/2-Y,1/2+Z REMARK 290 7555 1/2-X,1/2+Y,1/2-Z REMARK 290 8555 1/2+X,1/2-Y,-Z REMARK 290 REMARK 290 WHERE NNN -> OPERATOR NUMBER REMARK 290 MMM -> TRANSLATION VECTOR REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY TRANSFORMATIONS REMARK 290 THE FOLLOWING TRANSFORMATIONS OPERATE ON THE ATOM/HETATM REMARK 290 RECORDS IN THIS ENTRY TO PRODUCE CRYSTALLOGRAPHICALLY REMARK 290 RELATED MOLECULES. REMARK 290 SMTRY1 1 1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 1 0.000000 1.000000 0.000000 0.00000 REMARK 290 SMTRY3 1 0.000000 0.000000 1.000000 0.00000 REMARK 290 SMTRY1 2 -1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 2 0.000000 -1.000000 0.000000 0.00000 REMARK 290 SMTRY3 2 0.000000 0.000000 1.000000 60.03842 REMARK 290 SMTRY1 3 -1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 3 0.000000 1.000000 0.000000 0.00000 REMARK 290 SMTRY3 3 0.000000 0.000000 -1.000000 60.03842 REMARK 290 SMTRY1 4 1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 4 0.000000 -1.000000 0.000000 0.00000 REMARK 290 SMTRY3 4 0.000000 0.000000 -1.000000 0.00000 REMARK 290 SMTRY1 5 1.000000 0.000000 0.000000 19.37534 REMARK 290 SMTRY2 5 0.000000 1.000000 0.000000 37.60529 REMARK 290 SMTRY3 5 0.000000 0.000000 1.000000 0.00000 REMARK 290 SMTRY1 6 -1.000000 0.000000 0.000000 19.37534 REMARK 290 SMTRY2 6 0.000000 -1.000000 0.000000 37.60529 REMARK 290 SMTRY3 6 0.000000 0.000000 1.000000 60.03842 REMARK 290 SMTRY1 7 -1.000000 0.000000 0.000000 19.37534 REMARK 290 SMTRY2 7 0.000000 1.000000 0.000000 37.60529 REMARK 290 SMTRY3 7 0.000000 0.000000 -1.000000 60.03842 REMARK 290 SMTRY1 8 1.000000 0.000000 0.000000 19.37534 REMARK 290 SMTRY2 8 0.000000 -1.000000 0.000000 37.60529 REMARK 290 SMTRY3 8 0.000000 0.000000 -1.000000 0.00000 REMARK 290 REMARK 290 REMARK: NULL REMARK 375 REMARK 375 SPECIAL POSITION REMARK 375 CA 600 LIES ON A SPECIAL POSITION. REMARK 450 REMARK 450 SOURCE REMARK 450 GIFT OF THE LAB OF F. QUIOCHO REMARK 470 REMARK 470 MISSING ATOM REMARK 470 THE FOLLOWING RESIDUES HAVE MISSING ATOMS (M=MODEL NUMBER; REMARK 470 RES=RESIDUE NAME; C=CHAIN IDENTIFIER; SSEQ=SEQUENCE NUMBER; REMARK 470 I=INSERTION CODE): REMARK 470 M RES CSSEQI ATOMS REMARK 470 LYS A 77 CB CG CD CE NZ REMARK 470 LYS A 115 CB CG CD CE NZ REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: CLOSE CONTACTS REMARK 500 REMARK 500 THE FOLLOWING ATOMS THAT ARE RELATED BY CRYSTALLOGRAPHIC REMARK 500 SYMMETRY ARE IN CLOSE CONTACT. SOME OF THESE MAY BE ATOMS REMARK 500 LOCATED ON SPECIAL POSITIONS IN THE CELL. ATOMS WITH REMARK 500 NON-BLANK ALTERNATE LOCATION INDICATORS ARE NOT INCLUDED REMARK 500 IN THE CALCULATIONS. REMARK 500 REMARK 500 DISTANCE CUTOFF: REMARK 500 2.2 ANGSTROMS FOR CONTACTS NOT INVOLVING HYDROGEN ATOMS REMARK 500 1.6 ANGSTROMS FOR CONTACTS INVOLVING HYDROGEN ATOMS REMARK 500 REMARK 500 ATM1 RES C SSEQI ATM2 RES C SSEQI SSYMOP DISTANCE REMARK 500 CA CA 600 CA CA 600 4576 0.00 REMARK 850 REMARK 850 CORRECTION BEFORE RELEASE REMARK 850 ORIGINAL DEPOSITION REVISED PRIOR TO RELEASE REMARK 850 DATE REVISED: 23-SEP-1997 TRACKING NUMBER: T13047 REMARK 999 REMARK 999 SEQUENCE REMARK 999 1CM1 A SWS P02593 1 - 3 NOT IN ATOMS LIST REMARK 999 1CM1 A SWS P02593 147 - 148 NOT IN ATOMS LIST REMARK 999 1CM1 B SWS P11275 1 - 292 NOT IN ATOMS LIST REMARK 999 1CM1 B SWS P11275 311 - 478 NOT IN ATOMS LIST DBREF 1CM1 A 4 146 SWS P02593 CALM_HUMAN 4 146 DBREF 1CM1 B 293 310 SWS P11275 KCCA_RAT 293 310 SEQRES 1 A 148 ALA ASP GLN LEU THR GLU GLU GLN ILE ALA GLU PHE LYS SEQRES 2 A 148 GLU ALA PHE SER LEU PHE ASP LYS ASP GLY ASP GLY THR SEQRES 3 A 148 ILE THR THR LYS GLU LEU GLY THR VAL MET ARG SER LEU SEQRES 4 A 148 GLY GLN ASN PRO THR GLU ALA GLU LEU GLN ASP MET ILE SEQRES 5 A 148 ASN GLU VAL ASP ALA ASP GLY ASN GLY THR ILE ASP PHE SEQRES 6 A 148 PRO GLU PHE LEU THR MET MET ALA ARG LYS MET LYS ASP SEQRES 7 A 148 THR ASP SER GLU GLU GLU ILE ARG GLU ALA PHE ARG VAL SEQRES 8 A 148 PHE ASP LYS ASP GLY ASN GLY TYR ILE SER ALA ALA GLU SEQRES 9 A 148 LEU ARG HIS VAL MET THR ASN LEU GLY GLU LYS LEU THR SEQRES 10 A 148 ASP GLU GLU VAL ASP GLU MET ILE ARG GLU ALA ASP ILE SEQRES 11 A 148 ASP GLY ASP GLY GLN VAL ASN TYR GLU GLU PHE VAL GLN SEQRES 12 A 148 MET MET THR ALA LYS SEQRES 1 B 25 LEU LYS LYS PHE ASN ALA ARG ARG LYS LEU LYS GLY ALA SEQRES 2 B 25 ILE LEU THR THR MET LEU ALA THR ARG ASN PHE SER HET CA A 401 1 HET CA A 402 1 HET CA A 403 1 HET CA A 404 1 HET CA 600 1 HETNAM CA CALCIUM ION FORMUL 3 CA 5(CA1 2+) FORMUL 4 HOH *58(H2 O1) HELIX 1 1 GLU A 6 PHE A 19 1 14 HELIX 2 2 THR A 29 SER A 38 1 10 HELIX 3 3 GLU A 45 VAL A 55 1 11 HELIX 4 4 PHE A 65 ALA A 73 1 9 HELIX 5 5 GLU A 84 PHE A 92 1 9 HELIX 6 6 ALA A 102 ASN A 111 1 10 HELIX 7 7 ASP A 118 ALA A 128 1 11 HELIX 8 8 TYR A 138 MET A 144 1 7 HELIX 9 9 ALA B 295 LEU B 308 1 14 CRYST1 38.751 75.209 120.073 90.00 90.00 90.00 C 2 2 21 8 ORIGX1 1.000000 0.000000 0.000000 0.00000 ORIGX2 0.000000 1.000000 0.000000 0.00000 ORIGX3 0.000000 0.000000 1.000000 0.00000 SCALE1 0.025806 0.000000 0.000000 0.00000 SCALE2 0.000000 0.013296 0.000000 0.00000 SCALE3 0.000000 0.000000 0.008328 0.00000 (ATOM LINES ARE NOT SHOWN.) END