HEADER CALCIUM-REGULATED MUSCLE CONTRACTION 04-MAR-98 1TCF TITLE CRYSTAL STRUCTURE OF CALCIUM-SATURATED RABBIT SKELETAL TITLE 2 TROPONIN C COMPND MOL_ID: 1; COMPND 2 MOLECULE: TROPONIN C; COMPND 3 CHAIN: NULL; COMPND 4 ENGINEERED: YES; COMPND 5 MUTATION: INITIATOR MET, C98L SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: ORYCTOLAGUS CUNICULUS; SOURCE 3 ORGANISM_COMMON: RABBIT; SOURCE 4 EXPRESSION_SYSTEM: ESCHERICHIA COLI KEYWDS MUSCLE CONTRACTION, CALCIUM-BINDING, TROPONIN, E-F HAND, KEYWDS 2 OPEN CONFORMATION REGULATORY DOMAIN, CALCIUM-REGULATED KEYWDS 3 MUSCLE CONTRACTION EXPDTA X-RAY DIFFRACTION AUTHOR J.SOMAN,G.N.PHILLIPS JUNIOR REVDAT 1 27-MAY-98 1TCF 0 REMARK 1 REMARK 2 REMARK 2 RESOLUTION. 1.90 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) : 1.90 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 50.00 REMARK 3 DATA CUTOFF (SIGMA(F)) : 4.0 REMARK 3 DATA CUTOFF HIGH (ABS(F)) : 10000000.00 REMARK 3 DATA CUTOFF LOW (ABS(F)) : 0.001000 REMARK 3 COMPLETENESS (WORKING+TEST) (%) : 68.0 REMARK 3 NUMBER OF REFLECTIONS : 9587 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) : 0.278 REMARK 3 FREE R VALUE : 0.358 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 4.8 REMARK 3 FREE R VALUE TEST SET COUNT : 463 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 : 6 REMARK 3 BIN RESOLUTION RANGE HIGH (A) : 1.90 REMARK 3 BIN RESOLUTION RANGE LOW (A) : 2.02 REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 57.7 REMARK 3 REFLECTIONS IN BIN (WORKING SET) : 1286 REMARK 3 BIN R VALUE (WORKING SET) : 0.353 REMARK 3 BIN FREE R VALUE : 0.438 REMARK 3 BIN FREE R VALUE TEST SET SIZE (%) : 5.1 REMARK 3 BIN FREE R VALUE TEST SET COUNT : 69 REMARK 3 ESTIMATED ERROR OF BIN FREE R VALUE : 0.053 REMARK 3 REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT. REMARK 3 PROTEIN ATOMS : 1234 REMARK 3 NUCLEIC ACID ATOMS : 0 REMARK 3 HETEROGEN ATOMS : 5 REMARK 3 SOLVENT ATOMS : 197 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : 12.6 REMARK 3 MEAN B VALUE (OVERALL, A**2) : 22.0 REMARK 3 OVERALL ANISOTROPIC B VALUE. REMARK 3 B11 (A**2) : 1.38 REMARK 3 B22 (A**2) : -1.75 REMARK 3 B33 (A**2) : 0.37 REMARK 3 B12 (A**2) : 0.00 REMARK 3 B13 (A**2) : -0.57 REMARK 3 B23 (A**2) : 0.00 REMARK 3 REMARK 3 ESTIMATED COORDINATE ERROR. REMARK 3 ESD FROM LUZZATI PLOT (A) : 0.30 REMARK 3 ESD FROM SIGMAA (A) : 0.26 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) : 0.42 REMARK 3 ESD FROM C-V SIGMAA (A) : 0.39 REMARK 3 REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES. REMARK 3 BOND LENGTHS (A) : 0.016 REMARK 3 BOND ANGLES (DEGREES) : 1.9 REMARK 3 DIHEDRAL ANGLES (DEGREES) : 23.1 REMARK 3 IMPROPER ANGLES (DEGREES) : 1.06 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) : 2.31 ; 1.50 REMARK 3 MAIN-CHAIN ANGLE (A**2) : 3.25 ; 2.00 REMARK 3 SIDE-CHAIN BOND (A**2) : 3.86 ; 2.00 REMARK 3 SIDE-CHAIN ANGLE (A**2) : 5.82 ; 2.50 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 : NULL REMARK 3 TOPOLOGY FILE 1 : TOPHCSDX.PRO REMARK 3 TOPOLOGY FILE 2 : NULL REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: REMARK 3 KSOL : 0.32 REMARK 3 BSOL : 64.73 REMARK 6 REMARK 6 THE FIRST N-TERMINAL RESIDUE AND THE LAST TWO C-TERMINAL REMARK 6 RESIDUES ARE NOT IN THE ATOM LIST. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : NULL REMARK 200 TEMPERATURE (KELVIN) : 100 REMARK 200 PH : 5.6 REMARK 200 NUMBER OF CRYSTALS USED : 2 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : N REMARK 200 RADIATION SOURCE : CU REMARK 200 BEAMLINE : NULL REMARK 200 X-RAY GENERATOR MODEL : SIEMENS REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 1.54 REMARK 200 MONOCHROMATOR : NULL REMARK 200 OPTICS : NULL REMARK 200 REMARK 200 DETECTOR TYPE : R-AXIS IIC REMARK 200 DETECTOR MANUFACTURER : RIGAKU REMARK 200 INTENSITY-INTEGRATION SOFTWARE : DENZO REMARK 200 DATA SCALING SOFTWARE : SCALEPACK REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 10911 REMARK 200 RESOLUTION RANGE HIGH (A) : 1.90 REMARK 200 RESOLUTION RANGE LOW (A) : 50.0 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : 4.0 REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 77.5 REMARK 200 DATA REDUNDANCY : 4.0 REMARK 200 R MERGE (I) : 0.087 REMARK 200 R SYM (I) : NULL REMARK 200 <I/SIGMA(I)> FOR THE DATA SET : NULL 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: NULL REMARK 200 SOFTWARE USED: NULL REMARK 200 STARTING MODEL: NULL REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 52.5 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.6 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 40-50% MPD, 50MM SODIUM ACETATE REMARK 280 5MM CACL2, PH 5.6. REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 1 21 1 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 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 29.79915 REMARK 290 SMTRY3 2 0.000000 0.000000 -1.000000 0.00000 REMARK 290 REMARK 290 REMARK: NULL 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. AN ATOM LOCATED WITHIN 0.15 REMARK 500 ANGSTROMS OF A SYMMETRY RELATED ATOM IS ASSUMED TO BE ON A REMARK 500 SPECIAL POSITION AND IS, THEREFORE, LISTED IN REMARK 375 REMARK 500 INSTEAD OF REMARK 500. ATOMS WITH NON-BLANK ALTERNATE REMARK 500 LOCATION INDICATORS ARE NOT INCLUDED 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 NH2 ARG 100 O ASP 24 2656 2.05 REMARK 999 REMARK 999 SEQUENCE REMARK 999 1TCF SWS P02586 1 - 1 NOT IN ATOMS LIST REMARK 999 1TCF SWS P02586 158 - 159 NOT IN ATOMS LIST DBREF 1TCF 2 157 SWS P02586 TPCS_RABIT 2 157 SEQADV 1TCF LEU 98 SWS P02586 CYS 98 ENGINEERED SEQRES 1 159 THR ASP GLN GLN ALA GLU ALA ARG SER TYR LEU SER GLU SEQRES 2 159 GLU MET ILE ALA GLU PHE LYS ALA ALA PHE ASP MET PHE SEQRES 3 159 ASP ALA ASP GLY GLY GLY ASP ILE SER VAL LYS GLU LEU SEQRES 4 159 GLY THR VAL MET ARG MET LEU GLY GLN THR PRO THR LYS SEQRES 5 159 GLU GLU LEU ASP ALA ILE ILE GLU GLU VAL ASP GLU ASP SEQRES 6 159 GLY SER GLY THR ILE ASP PHE GLU GLU PHE LEU VAL MET SEQRES 7 159 MET VAL ARG GLN MET LYS GLU ASP ALA LYS GLY LYS SER SEQRES 8 159 GLU GLU GLU LEU ALA GLU LEU PHE ARG ILE PHE ASP ARG SEQRES 9 159 ASN ALA ASP GLY TYR ILE ASP ALA GLU GLU LEU ALA GLU SEQRES 10 159 ILE PHE ARG ALA SER GLY GLU HIS VAL THR ASP GLU GLU SEQRES 11 159 ILE GLU SER LEU MET LYS ASP GLY ASP LYS ASN ASN ASP SEQRES 12 159 GLY ARG ILE ASP PHE ASP GLU PHE LEU LYS MET MET GLU SEQRES 13 159 GLY VAL GLN HET CA 160 1 HET CA 161 1 HET CA 162 1 HET CA 163 1 HET CA 164 1 HETNAM CA CALCIUM ION FORMUL 2 CA 5(CA1 2+) FORMUL 3 HOH *197(H2 O1) HELIX 1 1 ALA 5 TYR 10 1 6 HELIX 2 2 GLU 13 PHE 26 1 14 HELIX 3 3 VAL 36 MET 45 1 10 HELIX 4 4 LYS 52 VAL 62 1 11 HELIX 5 5 PHE 72 MET 83 1 12 HELIX 6 6 GLU 93 PHE 102 1 10 HELIX 7 7 ALA 112 SER 122 1 11 HELIX 8 8 ASP 128 GLY 138 1 11 HELIX 9 9 PHE 148 GLU 156 1 9 LINK CA CA 160 OD1 ASP 27 LINK CA CA 160 OD1 ASP 29 LINK CA CA 160 O ASP 33 LINK CA CA 161 OD1 ASP 63 LINK CA CA 161 OD1 ASP 65 LINK CA CA 161 OG SER 67 LINK CA CA 161 OE1 GLU 74 LINK CA CA 162 OD1 ASP 103 LINK CA CA 162 OD1 ASP 107 LINK CA CA 162 OE1 GLU 114 LINK CA CA 162 O TYR 109 LINK CA CA 163 OD1 ASP 139 LINK CA CA 163 OD1 ASP 143 LINK CA CA 163 OE2 GLU 150 LINK CA CA 164 OE1 GLU 14 LINK CA CA 164 OE2 GLU 14 CRYST1 32.100 59.600 48.000 90.00 101.10 90.00 P 1 21 1 2 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 0.031153 0.000000 0.006112 0.00000 SCALE2 0.000000 0.016779 0.000000 0.00000 SCALE3 0.000000 0.000000 0.021231 0.00000 (ATOM LINES ARE NOT SHOWN.) END