HEADER CALCIUM-BINDING PROTEIN 10-APR-97 1AHR TITLE CALMODULIN MUTANT WITH A TWO RESIDUE DELETION IN THE TITLE 2 CENTRAL HELIX COMPND MOL_ID: 1; COMPND 2 MOLECULE: CALMODULIN; COMPND 3 CHAIN: NULL; COMPND 4 ENGINEERED: YES; COMPND 5 MUTATION: DEL(T79), DEL(D80) SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: GALLUS GALLUS; SOURCE 3 ORGANISM_COMMON: CHICKEN; SOURCE 4 EXPRESSION_SYSTEM: ESCHERICHIA COLI KEYWDS CALMODULIN, CALCIUM-BINDING PROTEIN EXPDTA X-RAY DIFFRACTION AUTHOR L.TABERNERO,J.SACK REVDAT 1 16-JUN-97 1AHR 0 JRNL AUTH L.TABERNERO,D.A.TAYLOR,R.J.CHANDROSS, JRNL AUTH 2 M.F.A.VANBERKUM,A.R.MEANS,F.A.QUIOCHO,J.S.SACK JRNL TITL THE STRUCTURE OF A CALMODULIN MUTANT WITH A JRNL TITL 2 DELETION IN THE CENTRAL HELIX SHOWS HOW CHANGES IN JRNL TITL 3 THE CONFORMATION OF CALMODULIN EFFECT ITS FUNCTION JRNL REF TO BE PUBLISHED JRNL REFN 0353 REMARK 1 REMARK 2 REMARK 2 RESOLUTION. 1.8 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : X-PLOR 3.1 REMARK 3 AUTHORS : BRUNGER REMARK 3 REMARK 3 DATA USED IN REFINEMENT. REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 1.8 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 8.0 REMARK 3 DATA CUTOFF (SIGMA(F)) : 2.0 REMARK 3 DATA CUTOFF HIGH (ABS(F)) : NULL REMARK 3 DATA CUTOFF LOW (ABS(F)) : NULL REMARK 3 COMPLETENESS (WORKING+TEST) (%) : NULL REMARK 3 NUMBER OF REFLECTIONS : NULL REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 CROSS-VALIDATION METHOD : NULL REMARK 3 FREE R VALUE TEST SET SELECTION : NULL REMARK 3 R VALUE (WORKING SET) : 0.211 REMARK 3 FREE R VALUE : NULL REMARK 3 FREE R VALUE TEST SET SIZE (%) : NULL REMARK 3 FREE R VALUE TEST SET COUNT : NULL REMARK 3 ESTIMATED ERROR OF FREE R VALUE : NULL REMARK 3 REMARK 3 FIT IN THE HIGHEST RESOLUTION BIN. REMARK 3 TOTAL NUMBER OF BINS USED : NULL REMARK 3 BIN RESOLUTION RANGE HIGH (A) : NULL REMARK 3 BIN RESOLUTION RANGE LOW (A) : NULL REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : NULL REMARK 3 REFLECTIONS IN BIN (WORKING SET) : NULL REMARK 3 BIN R VALUE (WORKING SET) : NULL REMARK 3 BIN FREE R VALUE : NULL REMARK 3 BIN FREE R VALUE TEST SET SIZE (%) : NULL REMARK 3 BIN FREE R VALUE TEST SET COUNT : NULL REMARK 3 ESTIMATED ERROR OF BIN FREE R VALUE : NULL REMARK 3 REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT. REMARK 3 PROTEIN ATOMS : 1149 REMARK 3 NUCLEIC ACID ATOMS : 0 REMARK 3 HETEROGEN ATOMS : 4 REMARK 3 SOLVENT ATOMS : 113 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : NULL REMARK 3 MEAN B VALUE (OVERALL, A**2) : NULL 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) : NULL REMARK 3 ESD FROM SIGMAA (A) : NULL REMARK 3 LOW RESOLUTION CUTOFF (A) : NULL REMARK 3 REMARK 3 CROSS-VALIDATED ESTIMATED COORDINATE ERROR. REMARK 3 ESD FROM C-V LUZZATI PLOT (A) : NULL REMARK 3 ESD FROM C-V SIGMAA (A) : NULL REMARK 3 REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES. REMARK 3 BOND LENGTHS (A) : 0.0150 REMARK 3 BOND ANGLES (DEGREES) : 1.8 REMARK 3 DIHEDRAL ANGLES (DEGREES) : NULL REMARK 3 IMPROPER ANGLES (DEGREES) : NULL REMARK 3 REMARK 3 ISOTROPIC THERMAL MODEL : NULL 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 : 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 : NULL REMARK 3 PARAMETER FILE 2 : NULL REMARK 3 TOPOLOGY FILE 1 : NULL REMARK 3 TOPOLOGY FILE 2 : NULL REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: NULL REMARK 4 REMARK 4 1AHR COMPLIES WITH FORMAT V. 2.2, 16-DEC-1996 REMARK 6 REMARK 6 SEQUENCE AND NUMBERING IS AS IN WILD TYPE CALMODULIN REMARK 6 (PDB ENTRY 4CLN) WITH RESIDUES THR 79 AND ASP 80 DELETED. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : DEC-1988 REMARK 200 TEMPERATURE (KELVIN) : 300 REMARK 200 PH : 4.25 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 RU-200 REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 1.5418 REMARK 200 MONOCHROMATOR : GRAPHITE REMARK 200 OPTICS : COLLIMATOR REMARK 200 REMARK 200 DETECTOR TYPE : MULTIWIRE AREA DETECTOR REMARK 200 DETECTOR MANUFACTURER : ADSC REMARK 200 INTENSITY-INTEGRATION SOFTWARE : UCSD SOFTWARE REMARK 200 DATA SCALING SOFTWARE : ROCKS REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 14407 REMARK 200 RESOLUTION RANGE HIGH (A) : 1.8 REMARK 200 RESOLUTION RANGE LOW (A) : NULL REMARK 200 REJECTION CRITERIA (SIGMA(I)) : 2. REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 67.3 REMARK 200 DATA REDUNDANCY : 2.4 REMARK 200 R MERGE (I) : NULL REMARK 200 R SYM (I) : 0.501 REMARK 200 <I/SIGMA(I)> FOR THE DATA SET : 8.68 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : NULL REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : NULL REMARK 200 COMPLETENESS FOR SHELL (%) : NULL REMARK 200 DATA REDUNDANCY IN SHELL : NULL REMARK 200 R MERGE FOR SHELL (I) : NULL REMARK 200 R SYM FOR SHELL (I) : NULL 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: MERLOT REMARK 200 STARTING MODEL: PDB ENTRY 4CLN REMARK 200 REMARK 200 REMARK: SEPARATE ROTATION SEARCH FOR EACH DOMAIN REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): NULL REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.32 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 28-33% MPD, 15% ETHANOL, 5MM REMARK 280 CACL, 20MM NA ACETATE, PH 4.25 REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 21 21 21 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 1/2-X,-Y,1/2+Z REMARK 290 3555 -X,1/2+Y,1/2-Z REMARK 290 4555 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 13.54500 REMARK 290 SMTRY2 2 0.000000 -1.000000 0.000000 0.00000 REMARK 290 SMTRY3 2 0.000000 0.000000 1.000000 50.44390 REMARK 290 SMTRY1 3 -1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 3 0.000000 1.000000 0.000000 27.75465 REMARK 290 SMTRY3 3 0.000000 0.000000 -1.000000 50.44390 REMARK 290 SMTRY1 4 1.000000 0.000000 0.000000 13.54500 REMARK 290 SMTRY2 4 0.000000 -1.000000 0.000000 27.75465 REMARK 290 SMTRY3 4 0.000000 0.000000 -1.000000 0.00000 REMARK 290 REMARK 290 REMARK: NULL REMARK 525 REMARK 525 SOLVENT REMARK 525 THE FOLLOWING SOLVENT MOLECULES LIE FARTHER THAN EXPECTED REMARK 525 FROM THE PROTEIN OR NUCLEIC ACID MOLECULE AND MAY BE REMARK 525 ASSOCIATED WITH A SYMMETRY RELATED MOLECULE (M=MODEL REMARK 525 NUMBER; RES=RESIDUE NAME; C=CHAIN IDENTIFIER; SSEQ=SEQUENCE REMARK 525 NUMBER; I=INSERTION CODE): REMARK 525 REMARK 525 M RES CSSEQI REMARK 525 0 HOH 273 DISTANCE = 5.36 ANGSTROMS REMARK 525 0 HOH 293 DISTANCE = 5.18 ANGSTROMS DBREF 1AHR 1 148 SWS P07181 CALM_DROME 1 148 SEQADV 1AHR SWS P07181 THR 79 ENGINEERED DELETION SEQADV 1AHR SWS P07181 ASP 80 ENGINEERED DELETION SEQRES 1 146 ALA ASP GLN LEU THR GLU GLU GLN ILE ALA GLU PHE LYS SEQRES 2 146 GLU ALA PHE SER LEU PHE ASP LYS ASP GLY ASP GLY THR SEQRES 3 146 ILE THR THR LYS GLU LEU GLY THR VAL MET ARG SER LEU SEQRES 4 146 GLY GLN ASN PRO THR GLU ALA GLU LEU GLN ASP MET ILE SEQRES 5 146 ASN GLU VAL ASP ALA ASP GLY ASN GLY THR ILE ASP PHE SEQRES 6 146 PRO GLU PHE LEU THR MET MET ALA ARG LYS MET LYS ASP SEQRES 7 146 SER GLU GLU GLU ILE ARG GLU ALA PHE ARG VAL PHE ASP SEQRES 8 146 LYS ASP GLY ASN GLY PHE ILE SER ALA ALA GLU LEU ARG SEQRES 9 146 HIS VAL MET THR ASN LEU GLY GLU LYS LEU THR ASP GLU SEQRES 10 146 GLU VAL ASP GLU MET ILE ARG GLU ALA ASP ILE ASP GLY SEQRES 11 146 ASP GLY GLN VAL ASN TYR GLU GLU PHE VAL THR MET MET SEQRES 12 146 THR SER LYS HET CA 149 1 HET CA 150 1 HET CA 151 1 HET CA 152 1 HETNAM CA CALCIUM ION FORMUL 2 CA 4(CA1 2+) FORMUL 3 HOH *113(H2 O1) HELIX 1 1 GLN 8 PHE 19 1 12 HELIX 2 2 THR 29 SER 38 1 10 HELIX 3 3 GLU 45 VAL 55 1 11 HELIX 4 4 PHE 65 PHE 92 1 26 HELIX 5 5 ALA 102 LEU 112 1 11 HELIX 6 6 ASP 118 GLU 127 1 10 HELIX 7 7 TYR 138 MET 144 1 7 SHEET 1 A 2 PHE 99 SER 101 0 SHEET 2 A 2 GLN 135 ASN 137 -1 N VAL 136 O ILE 100 LINK CA CA 149 OD1 ASP 20 LINK CA CA 149 O THR 26 LINK CA CA 149 OE1 GLU 31 LINK CA CA 150 OD1 ASP 56 LINK CA CA 150 OD1 ASN 60 LINK CA CA 150 O THR 62 LINK CA CA 150 OE2 GLU 67 LINK CA CA 151 OD1 ASP 93 LINK CA CA 151 OD1 ASN 97 LINK CA CA 151 O PHE 99 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 OE2 GLU 140 CRYST1 27.090 55.510 100.890 90.00 90.00 90.00 P 21 21 21 4 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 0.036914 0.000000 0.000000 0.00000 SCALE2 0.000000 0.018015 0.000000 0.00000 SCALE3 0.000000 0.000000 0.009912 0.00000 (ATOM LINES ARE NOT SHOWN.) END