HEADER CALCIUM-BINDING PROTEIN 19-JAN-98 1A29 TITLE CALMODULIN COMPLEXED WITH TRIFLUOPERAZINE (1:2 COMPLEX) COMPND MOL_ID: 1; COMPND 2 MOLECULE: CALMODULIN; COMPND 3 CHAIN: NULL; COMPND 4 BIOLOGICAL_UNIT: MONOMER SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: BOS TAURUS; SOURCE 3 ORGANISM_COMMON: BOVINE; SOURCE 4 ORGAN: BRAIN; SOURCE 5 CELLULAR_LOCATION: CYTOPLASM KEYWDS CALCIUM-BINDING PROTEIN EXPDTA X-RAY DIFFRACTION AUTHOR ZS.BOCSKEI,V.HARMAT,B.G.VERTESSY,J.OVADI,G.NARAY-SZABO REVDAT 1 16-SEP-98 1A29 0 JRNL AUTH M.VANDONSELAAR,R.A.HICKIE,J.W.QUAIL,L.T.DELBAERE JRNL TITL TRIFLUOPERAZINE-INDUCED CONFORMATIONAL CHANGE IN JRNL TITL 2 CA(2+)-CALMODULIN JRNL REF NAT.STRUCT.BIOL. V. 1 795 1994 JRNL REFN ASTM NSBIEW US ISSN 1072-8368 2024 REMARK 1 REMARK 1 REFERENCE 1 REMARK 1 AUTH W.J.COOK,L.J.WALTER,M.R.WALTER REMARK 1 TITL DRUG BINDING BY CALMODULIN: CRYSTAL STRUCTURE OF A REMARK 1 TITL 2 CALMODULIN-TRIFLUOPERAZINE COMPLEX REMARK 1 REF BIOCHEMISTRY V. 33 15259 1994 REMARK 1 REFN ASTM BICHAW US ISSN 0006-2960 0033 REMARK 2 REMARK 2 RESOLUTION. 2.74 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) : 2.74 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 59.19 REMARK 3 DATA CUTOFF (SIGMA(F)) : 2 REMARK 3 DATA CUTOFF HIGH (ABS(F)) : 1000000 REMARK 3 DATA CUTOFF LOW (ABS(F)) : 0.001 REMARK 3 COMPLETENESS (WORKING+TEST) (%) : 96.46 REMARK 3 NUMBER OF REFLECTIONS : 4768 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) : 0.1971 REMARK 3 FREE R VALUE : 0.2654 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 4.8 REMARK 3 FREE R VALUE TEST SET COUNT : 230 REMARK 3 ESTIMATED ERROR OF FREE R VALUE : 0.017 REMARK 3 REMARK 3 FIT IN THE HIGHEST RESOLUTION BIN. REMARK 3 TOTAL NUMBER OF BINS USED : 8 REMARK 3 BIN RESOLUTION RANGE HIGH (A) : 2.74 REMARK 3 BIN RESOLUTION RANGE LOW (A) : 2.82 REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 57.3 REMARK 3 REFLECTIONS IN BIN (WORKING SET) : 233 REMARK 3 BIN R VALUE (WORKING SET) : 0.1661 REMARK 3 BIN FREE R VALUE : 0.3395 REMARK 3 BIN FREE R VALUE TEST SET SIZE (%) : 4.9 REMARK 3 BIN FREE R VALUE TEST SET COUNT : 12 REMARK 3 ESTIMATED ERROR OF BIN FREE R VALUE : 0.098 REMARK 3 REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT. REMARK 3 PROTEIN ATOMS : 1087 REMARK 3 NUCLEIC ACID ATOMS : 0 REMARK 3 HETEROGEN ATOMS : 60 REMARK 3 SOLVENT ATOMS : 0 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : 32.0 REMARK 3 MEAN B VALUE (OVERALL, A**2) : 30.3 REMARK 3 OVERALL ANISOTROPIC B VALUE. REMARK 3 B11 (A**2) : -9.975 REMARK 3 B22 (A**2) : -9.975 REMARK 3 B33 (A**2) : -7.101 REMARK 3 B12 (A**2) : -8.454 REMARK 3 B13 (A**2) : 0.000 REMARK 3 B23 (A**2) : 0.000 REMARK 3 REMARK 3 ESTIMATED COORDINATE ERROR. REMARK 3 ESD FROM LUZZATI PLOT (A) : 0.27 REMARK 3 ESD FROM SIGMAA (A) : 0.12 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) : 0.40 REMARK 3 ESD FROM C-V SIGMAA (A) : 0.22 REMARK 3 REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES. REMARK 3 BOND LENGTHS (A) : 0.006 REMARK 3 BOND ANGLES (DEGREES) : 1.08 REMARK 3 DIHEDRAL ANGLES (DEGREES) : 21.5 REMARK 3 IMPROPER ANGLES (DEGREES) : 1.10 REMARK 3 REMARK 3 ISOTROPIC THERMAL MODEL : GROUPED REMARK 3 REMARK 3 ISOTROPIC THERMAL FACTOR RESTRAINTS. RMS SIGMA REMARK 3 MAIN-CHAIN BOND (A**2) : NULL ; NULL REMARK 3 MAIN-CHAIN ANGLE (A**2) : NULL ; NULL REMARK 3 SIDE-CHAIN BOND (A**2) : NULL ; NULL REMARK 3 SIDE-CHAIN ANGLE (A**2) : NULL ; NULL REMARK 3 REMARK 3 NCS MODEL : NONE 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 : PARHCSDX.PRO REMARK 3 PARAMETER FILE 2 : CA.PAR REMARK 3 PARAMETER FILE 3 : TFP.PAR REMARK 3 TOPOLOGY FILE 1 : TOPHCSDX.PRO REMARK 3 TOPOLOGY FILE 2 : CA.TOP REMARK 3 TOPOLOGY FILE 3 : TFP.TOP REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: NULL REMARK 4 REMARK 4 1A29 COMPLIES WITH FORMAT V. 2.2, 16-DEC-1996 REMARK 6 REMARK 6 THE DISORDERED SIDE CHAIN ATOMS OF GLU 6, GLU 7, ASN 53, REMARK 6 ARG 74, LYS 75, MET 76, LYS 77, ASP 78, THR 79, ASP 80, REMARK 6 GLU 84, GLU 87, LYS 94, LYS 115 AS WELL AS THE DISORDERED REMARK 6 RESIDUES ALA 1, ASP 2, ALA 147, LYS 148 ARE NOT IN ATOMS REMARK 6 LIST. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : FEB-1995 REMARK 200 TEMPERATURE (KELVIN) : 293 REMARK 200 PH : 5.0 REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : N REMARK 200 RADIATION SOURCE : NULL REMARK 200 BEAMLINE : NULL REMARK 200 X-RAY GENERATOR MODEL : RIGAKU RU200 REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 1.5418 REMARK 200 MONOCHROMATOR : GRAPHITE REMARK 200 OPTICS : NORMAL FOCUS REMARK 200 REMARK 200 DETECTOR TYPE : IMAGE PLATE REMARK 200 DETECTOR MANUFACTURER : RIGAKU REMARK 200 INTENSITY-INTEGRATION SOFTWARE : BIOTEX REMARK 200 DATA SCALING SOFTWARE : BIOTEX REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 17223 REMARK 200 RESOLUTION RANGE HIGH (A) : 2.74 REMARK 200 RESOLUTION RANGE LOW (A) : 87.7 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : 1.0 REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 98.0 REMARK 200 DATA REDUNDANCY : 1.92 REMARK 200 R MERGE (I) : 0.086 REMARK 200 R SYM (I) : 0.0737 REMARK 200 <I/SIGMA(I)> FOR THE DATA SET : 11.3 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 2.74 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 2.86 REMARK 200 COMPLETENESS FOR SHELL (%) : 98.2 REMARK 200 DATA REDUNDANCY IN SHELL : 1.90 REMARK 200 R MERGE FOR SHELL (I) : 0.092 REMARK 200 R SYM FOR SHELL (I) : 0.205 REMARK 200 <I/SIGMA(I)> FOR SHELL : 1.84 REMARK 200 REMARK 200 DIFFRACTION PROTOCOL: NULL REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: MOLECULAR REMARK 200 REPLACEMENT REMARK 200 SOFTWARE USED: AMORE REMARK 200 STARTING MODEL: PDB ENTRY 1LIN REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 52 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.5 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: PROTEIN WAS CRYSTALLIZED BY REMARK 280 SITTING DROP TECHNIQUE IN THE COLD ROOM (8 DEGREES C +/- 2 REMARK 280 DEGREES C) BY MIXING 4 MICROLITERS OF 1 MM PROTEIN REMARK 280 CONTAINING 1.2-1.5 MM TFP IN 5 MM CACL2 WITH 4 MICROLITERS REMARK 280 OF THE RESERVOIR SOLUTION (1 ML 10 MM SODIUM CACODYLATE/HCL REMARK 280 BUFFER, PH 5.2-5.6 WITH 10 MM CACL2, 25-30 % (W/V) REMARK 280 POLYETHYLENE GLYCOL 6000), CRYSTAL GROWTH TOOK 2-3 WEEKS. REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 32 2 1 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 -Y,X-Y,Z+2/3 REMARK 290 3555 Y-X,-X,Z+1/3 REMARK 290 4555 Y,X,-Z REMARK 290 5555 X-Y,-Y,1/3-Z REMARK 290 6555 -X,Y-X,2/3-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 -0.500000 -0.866015 0.000000 0.00000 REMARK 290 SMTRY2 2 0.866036 -0.500000 0.000000 0.00000 REMARK 290 SMTRY3 2 0.000000 0.000000 1.000000 118.37121 REMARK 290 SMTRY1 3 -0.500000 0.866015 0.000000 0.00000 REMARK 290 SMTRY2 3 -0.866036 -0.500000 0.000000 0.00000 REMARK 290 SMTRY3 3 0.000000 0.000000 1.000000 59.18561 REMARK 290 SMTRY1 4 -0.500000 0.866015 0.000000 0.00000 REMARK 290 SMTRY2 4 0.866036 0.500000 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 0.00000 REMARK 290 SMTRY2 5 0.000000 -1.000000 0.000000 0.00000 REMARK 290 SMTRY3 5 0.000000 0.000000 -1.000000 59.18561 REMARK 290 SMTRY1 6 -0.500000 -0.866015 0.000000 0.00000 REMARK 290 SMTRY2 6 -0.866036 0.500000 0.000000 0.00000 REMARK 290 SMTRY3 6 0.000000 0.000000 -1.000000 118.37121 REMARK 290 REMARK 290 REMARK: NULL 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 GLU 6 CG CD OE1 OE2 REMARK 470 GLU 7 CG CD OE1 OE2 REMARK 470 ASN 53 CG OD1 ND2 REMARK 470 ARG 74 CG CD NE CZ NH1 NH2 REMARK 470 LYS 75 CG CD CE NZ REMARK 470 MET 76 CG SD CE REMARK 470 LYS 77 CG CD CE NZ REMARK 470 ASP 78 CG OD1 OD2 REMARK 470 THR 79 OG1 CG2 REMARK 470 ASP 80 CG OD1 OD2 REMARK 470 GLU 84 CG CD OE1 OE2 REMARK 470 GLU 87 CG CD OE1 OE2 REMARK 470 LYS 94 CG CD CE NZ REMARK 470 LYS 115 CG CD CE NZ REMARK 850 REMARK 850 CORRECTION BEFORE RELEASE REMARK 850 ORIGINAL DEPOSITION REVISED PRIOR TO RELEASE REMARK 850 DATE REVISED: 17-FEB-1998 TRACKING NUMBER: T14115 REMARK 999 REMARK 999 SEQUENCE REMARK 999 REFERENCE: THE SEQUENCE IS DESCRIBED IN WATTERSON ET AL. REMARK 999 J BIOL CHEM 1980 FEB 10;255(3):962-975. REMARK 999 1A29 SWS P02593 1 - 2 NOT IN ATOMS LIST REMARK 999 1A29 SWS P02593 147 - 148 NOT IN ATOMS LIST DBREF 1A29 3 146 SWS P02593 CALM_HUMAN 3 146 SEQRES 1 148 ALA ASP GLN LEU THR GLU GLU GLN ILE ALA GLU PHE LYS SEQRES 2 148 GLU ALA PHE SER LEU PHE ASP LYS ASP GLY ASP GLY THR SEQRES 3 148 ILE THR THR LYS GLU LEU GLY THR VAL MET ARG SER LEU SEQRES 4 148 GLY GLN ASN PRO THR GLU ALA GLU LEU GLN ASP MET ILE SEQRES 5 148 ASN GLU VAL ASP ALA ASP GLY ASN GLY THR ILE ASP PHE SEQRES 6 148 PRO GLU PHE LEU THR MET MET ALA ARG LYS MET LYS ASP SEQRES 7 148 THR ASP SER GLU GLU GLU ILE ARG GLU ALA PHE ARG VAL SEQRES 8 148 PHE ASP LYS ASP GLY ASN GLY TYR ILE SER ALA ALA GLU SEQRES 9 148 LEU ARG HIS VAL MET THR ASN LEU GLY GLU LYS LEU THR SEQRES 10 148 ASP GLU GLU VAL ASP GLU MET ILE ARG GLU ALA ASP ILE SEQRES 11 148 ASP GLY ASP GLY GLN VAL ASN TYR GLU GLU PHE VAL GLN SEQRES 12 148 MET MET THR ALA LYS HET CA 149 1 HET CA 150 1 HET CA 151 1 HET CA 152 1 HET TFP 153 28 HET TFP 154 28 HETNAM CA CALCIUM ION HETNAM TFP TRIFLUO-METHYL-PERAZINE FORMUL 2 CA 4(CA1 2+) FORMUL 3 TFP 2(C21 H24 N3 F3 S1) HELIX 1 1 GLU 6 PHE 19 1 14 HELIX 2 2 THR 29 SER 38 1 10 HELIX 3 3 GLU 45 VAL 55 1 11 HELIX 4 4 PHE 65 ALA 73 1 9 HELIX 5 5 GLU 82 PHE 92 1 11 HELIX 6 6 ALA 102 LEU 112 1 11 HELIX 7 7 ASP 118 ALA 128 1 11 HELIX 8 8 TYR 138 MET 144 1 7 LINK CA CA 149 OD1 ASP 20 LINK CA CA 149 OD1 ASP 22 LINK CA CA 149 OD1 ASP 24 LINK CA CA 149 O THR 26 LINK CA CA 149 OE1 GLU 31 LINK CA CA 149 OE2 GLU 31 LINK CA CA 150 OD1 ASP 56 LINK CA CA 150 OD1 ASP 58 LINK CA CA 150 OD1 ASN 60 LINK CA CA 150 O THR 62 LINK CA CA 150 OE1 GLU 67 LINK CA CA 150 OE2 GLU 67 LINK CA CA 151 OD1 ASP 93 LINK CA CA 151 OD1 ASP 95 LINK CA CA 151 OD1 ASN 97 LINK CA CA 151 O TYR 99 LINK CA CA 151 OE1 GLU 104 LINK CA CA 151 OE2 GLU 104 LINK CA CA 152 OD1 ASP 129 LINK CA CA 152 OD1 ASP 131 LINK CA CA 152 OD1 ASP 133 LINK CA CA 152 O GLN 135 LINK CA CA 152 OE1 GLU 140 LINK CA CA 152 OE2 GLU 140 CRYST1 40.750 40.750 177.570 90.00 90.00 120.00 P 32 2 1 6 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 0.024540 0.014168 0.000000 0.00000 SCALE2 0.000000 0.028336 0.000000 0.00000 SCALE3 0.000000 0.000000 0.005632 0.00000 (ATOM LINES ARE NOT SHOWN.) END