HEADER COMPLEX (SKELETAL MUSCLE/MUSCLE PROTEIN)13-JAN-98 1A2X TITLE COMPLEX OF TROPONIN C WITH A 47 RESIDUE (1-47) FRAGMENT OF TITLE 2 TROPONIN I COMPND MOL_ID: 1; COMPND 2 MOLECULE: TROPONIN C; COMPND 3 CHAIN: A; COMPND 4 ENGINEERED: YES; COMPND 5 BIOLOGICAL_UNIT: MONOMER WITHIN TERNARY COMPLEX WITH COMPND 6 TROPONIN I AND TROPONIN T; COMPND 7 MOL_ID: 2; COMPND 8 MOLECULE: TROPONIN I; COMPND 9 CHAIN: B; COMPND 10 FRAGMENT: RESIDUES 1 - 47; COMPND 11 ENGINEERED: YES; COMPND 12 BIOLOGICAL_UNIT: FULL LENGTH MONOMER WITHIN TERNARY COMPLEX COMPND 13 WITH TROPONIN C AND TROPONIN T SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: ORYCTOLAGUS CUNICULUS; SOURCE 3 ORGANISM_COMMON: RABBIT; SOURCE 4 TISSUE: FAST SKELETAL MUSCLE; SOURCE 5 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 6 EXPRESSION_SYSTEM_STRAIN: K; SOURCE 7 EXPRESSION_SYSTEM_CELLULAR_LOCATION: CYTOPLASM; SOURCE 8 EXPRESSION_SYSTEM_PLASMID: PMALC2; SOURCE 9 MOL_ID: 2; SOURCE 10 ORGANISM_SCIENTIFIC: ORYCTOLAGUS CUNICULUS; SOURCE 11 ORGANISM_COMMON: RABBIT; SOURCE 12 TISSUE: FAST SKELETAL MUSCLE; SOURCE 13 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 14 EXPRESSION_SYSTEM_STRAIN: K; SOURCE 15 EXPRESSION_SYSTEM_CELLULAR_LOCATION: INCLUSION BODY; SOURCE 16 EXPRESSION_SYSTEM_PLASMID: PTRC KEYWDS TROPONIN, MUSCLE CONTRACTION REGULATION, KEYWDS 2 COMPLEX (SKELETAL MUSCLE/MUSCLE PROTEIN) EXPDTA X-RAY DIFFRACTION AUTHOR D.G.VASSYLYEV,S.TAKEDA,S.WAKATSUKI,K.MAEDA,Y.MAEDA REVDAT 1 15-JUL-98 1A2X 0 JRNL AUTH D.G.VASSYLYEV,S.TAKEDA,S.WAKATSUKI,K.MAEDA,Y.MAEDA JRNL TITL CRYSTAL STRUCTURE OF TROPONIN C IN COMPLEX WITH JRNL TITL 2 TROPONIN I FRAGMENT AT 2.3-A RESOLUTION JRNL REF PROC.NAT.ACAD.SCI.USA V. 95 4847 1998 JRNL REFN ASTM PNASA6 US ISSN 0027-8424 0040 REMARK 1 REMARK 1 REFERENCE 1 REMARK 1 AUTH Y.SAIJO,S.TAKEDA,A.SCHERER,T.KOBAYASHI,Y.MAEDA, REMARK 1 AUTH 2 H.TANIGUCHI,M.YAO,S.WAKATSUKI REMARK 1 TITL PRODUCTION, CRYSTALLIZATION, AND PRELIMINARY X-RAY REMARK 1 TITL 2 ANALYSIS OF RABBIT SKELETAL MUSCLE TROPONIN COMPLEX REMARK 1 TITL 3 CONSISTING OF TROPONIN C AND FRAGMENT (1-47) OF REMARK 1 TITL 4 TROPONIN I REMARK 1 REF PROTEIN SCI. V. 6 916 1997 REMARK 1 REFN ASTM PRCIEI US ISSN 0961-8368 0795 REMARK 2 REMARK 2 RESOLUTION. 2.3 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) : 2.3 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 10.0 REMARK 3 DATA CUTOFF (SIGMA(F)) : 0. REMARK 3 DATA CUTOFF HIGH (ABS(F)) : 100000000000. REMARK 3 DATA CUTOFF LOW (ABS(F)) : 0. REMARK 3 COMPLETENESS (WORKING+TEST) (%) : 96.2 REMARK 3 NUMBER OF REFLECTIONS : 8736 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 CROSS-VALIDATION METHOD : NULL REMARK 3 FREE R VALUE TEST SET SELECTION : RANDOM REMARK 3 R VALUE (WORKING SET) : 0.222 REMARK 3 FREE R VALUE : 0.325 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 7. REMARK 3 FREE R VALUE TEST SET COUNT : 591 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 : 8 REMARK 3 BIN RESOLUTION RANGE HIGH (A) : 2.3 REMARK 3 BIN RESOLUTION RANGE LOW (A) : 2.4 REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 95.4 REMARK 3 REFLECTIONS IN BIN (WORKING SET) : 1000 REMARK 3 BIN R VALUE (WORKING SET) : 0.337 REMARK 3 BIN FREE R VALUE : 0.389 REMARK 3 BIN FREE R VALUE TEST SET SIZE (%) : 6.7 REMARK 3 BIN FREE R VALUE TEST SET COUNT : 58 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 : 1502 REMARK 3 NUCLEIC ACID ATOMS : 0 REMARK 3 HETEROGEN ATOMS : 2 REMARK 3 SOLVENT ATOMS : 89 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : 45. REMARK 3 MEAN B VALUE (OVERALL, A**2) : 41. 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) : 0.30 REMARK 3 ESD FROM SIGMAA (A) : 0.28 REMARK 3 LOW RESOLUTION CUTOFF (A) : 6. 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) : 0.01 REMARK 3 BOND ANGLES (DEGREES) : 1.28 REMARK 3 DIHEDRAL ANGLES (DEGREES) : 24.8 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) : 4.9 ; 1.5 REMARK 3 MAIN-CHAIN ANGLE (A**2) : 6.6 ; 2.0 REMARK 3 SIDE-CHAIN BOND (A**2) : 4.6 ; 2.0 REMARK 3 SIDE-CHAIN ANGLE (A**2) : 8.0 ; 2.5 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 : 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: NULL REMARK 4 REMARK 4 1A2X COMPLIES WITH FORMAT V. 2.2, 16-DEC-1996 REMARK 6 REMARK 6 IN CHAIN A, RESIDUE 1 IS DISORDERED. IN CHAIN B, RESIDUES REMARK 6 1, 2 AND 34 - 47 ARE DISORDERED. THESE RESIDUES ARE NOT REMARK 6 INCLUDED IN THE ENTRY. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : JUN-1997 REMARK 200 TEMPERATURE (KELVIN) : 100 REMARK 200 PH : 7.0 REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : Y REMARK 200 RADIATION SOURCE : ESRF REMARK 200 BEAMLINE : BM-14 REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 0.97 REMARK 200 MONOCHROMATOR : MONOCHROMATOR REMARK 200 OPTICS : NULL REMARK 200 REMARK 200 DETECTOR TYPE : 30 CM IMAGE PLATE REMARK 200 DETECTOR MANUFACTURER : MARRESEARCH REMARK 200 INTENSITY-INTEGRATION SOFTWARE : DENZO REMARK 200 DATA SCALING SOFTWARE : SCALEPACK REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 8870 REMARK 200 RESOLUTION RANGE HIGH (A) : 2.3 REMARK 200 RESOLUTION RANGE LOW (A) : 25. REMARK 200 REJECTION CRITERIA (SIGMA(I)) : 0. REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 96.3 REMARK 200 DATA REDUNDANCY : 4.5 REMARK 200 R MERGE (I) : 0.05 REMARK 200 R SYM (I) : 0.05 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) : 2.3 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 2.4 REMARK 200 COMPLETENESS FOR SHELL (%) : 94. REMARK 200 DATA REDUNDANCY IN SHELL : 4.3 REMARK 200 R MERGE FOR SHELL (I) : 0.30 REMARK 200 R SYM FOR SHELL (I) : NULL REMARK 200 <I/SIGMA(I)> FOR SHELL : 2.3 REMARK 200 REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: SIR/MAD REMARK 200 SOFTWARE USED: CCP4 REMARK 200 STARTING MODEL: NULL REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 35.0 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.3 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: HANGING-DROP VAPOR DIFFUSION REMARK 280 METHOD WAS USED AT 289K BY MIXING THE PROTEIN REMARK 280 SOLUTION CONTAINING 25-30MG/ML OF THE CI47 COMPLEX WITH A REMARK 280 RESERVOIR SOLUTION CONTAINING 1.5M SODIUM CITRATE, 0.1M REMARK 280 TRIS-HCL, PH 8.0, 15% TREHALOSE. 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 -0.866007 0.000000 0.00000 REMARK 290 SMTRY2 2 0.866044 -0.500000 0.000000 0.00000 REMARK 290 SMTRY3 2 0.000000 0.000000 1.000000 101.53315 REMARK 290 SMTRY1 3 -0.500000 0.866007 0.000000 0.00000 REMARK 290 SMTRY2 3 -0.866044 -0.500000 0.000000 0.00000 REMARK 290 SMTRY3 3 0.000000 0.000000 1.000000 50.76658 REMARK 290 SMTRY1 4 -0.500000 0.866007 0.000000 0.00000 REMARK 290 SMTRY2 4 0.866044 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 50.76658 REMARK 290 SMTRY1 6 -0.500000 -0.866007 0.000000 0.00000 REMARK 290 SMTRY2 6 -0.866044 0.500000 0.000000 0.00000 REMARK 290 SMTRY3 6 0.000000 0.000000 -1.000000 101.53315 REMARK 290 REMARK 290 REMARK: NULL REMARK 650 REMARK 650 HELIX REMARK 650 DETERMINATION METHOD: AUTHOR-DETERMINED REMARK 700 REMARK 700 SHEET REMARK 700 DETERMINATION METHOD: AUTHOR-DETERMINED REMARK 999 REMARK 999 SEQUENCE REMARK 999 1A2X A SWS P02586 1 - 1 NOT IN ATOMS LIST REMARK 999 1A2X B SWS P02643 1 - 2 NOT IN ATOMS LIST REMARK 999 1A2X B SWS P02643 34 - 181 NOT IN ATOMS LIST REMARK 999 REMARK 999 CHAIN B REPRESENTS A 47 RESIDUE FRAGMENT OF TROPONIN I. REMARK 999 RESIDUES 48 - 181 WERE NOT EXPRESSED AND ARE NOT INCLUDED REMARK 999 IN THIS ENTRY. DBREF 1A2X A 2 159 SWS P02586 TPCS_RABIT 2 159 DBREF 1A2X B 3 33 SWS P02643 TRIF_RABIT 3 33 SEQRES 1 A 159 THR ASP GLN GLN ALA GLU ALA ARG SER TYR LEU SER GLU SEQRES 2 A 159 GLU MET ILE ALA GLU PHE LYS ALA ALA PHE ASP MET PHE SEQRES 3 A 159 ASP ALA ASP GLY GLY GLY ASP ILE SER VAL LYS GLU LEU SEQRES 4 A 159 GLY THR VAL MET ARG MET LEU GLY GLN THR PRO THR LYS SEQRES 5 A 159 GLU GLU LEU ASP ALA ILE ILE GLU GLU VAL ASP GLU ASP SEQRES 6 A 159 GLY SER GLY THR ILE ASP PHE GLU GLU PHE LEU VAL MET SEQRES 7 A 159 MET VAL ARG GLN MET LYS GLU ASP ALA LYS GLY LYS SER SEQRES 8 A 159 GLU GLU GLU LEU ALA GLU CYS PHE ARG ILE PHE ASP ARG SEQRES 9 A 159 ASN ALA ASP GLY TYR ILE ASP ALA GLU GLU LEU ALA GLU SEQRES 10 A 159 ILE PHE ARG ALA SER GLY GLU HIS VAL THR ASP GLU GLU SEQRES 11 A 159 ILE GLU SER LEU MET LYS ASP GLY ASP LYS ASN ASN ASP SEQRES 12 A 159 GLY ARG ILE ASP PHE ASP GLU PHE LEU LYS MET MET GLU SEQRES 13 A 159 GLY VAL GLN SEQRES 1 B 47 GLY ASP GLU GLU LYS ARG ASN ARG ALA ILE THR ALA ARG SEQRES 2 B 47 ARG GLN HIS LEU LYS SER VAL MET LEU GLN ILE ALA ALA SEQRES 3 B 47 THR GLU LEU GLU LYS GLU GLU GLY ARG ARG GLU ALA GLU SEQRES 4 B 47 LYS GLN ASN TYR LEU ALA GLU HIS HET CA A 160 1 HET CA A 161 1 HETNAM CA CALCIUM ION FORMUL 3 CA 2(CA1 2+) FORMUL 4 HOH *89(H2 O1) HELIX 1 1 GLN A 4 TYR A 10 1 7 HELIX 2 2 GLU A 13 PHE A 26 1 14 HELIX 3 3 VAL A 36 MET A 45 1 10 HELIX 4 4 LYS A 52 VAL A 62 1 11 HELIX 5 5 PHE A 72 GLU A 85 1 14 HELIX 6 6 GLU A 92 PHE A 102 1 11 HELIX 7 7 ALA A 112 ILE A 118 1 7 HELIX 8 8 ASP A 128 GLY A 138 1 11 HELIX 9 9 PHE A 148 MET A 155 1 8 HELIX 10 10 LYS B 5 LYS B 31 1 27 SHEET 1 A 2 ASP A 33 SER A 35 0 SHEET 2 A 2 THR A 69 ASP A 71 -1 N ILE A 70 O ILE A 34 SHEET 1 B 2 TYR A 109 ASP A 111 0 SHEET 2 B 2 ARG A 145 ASP A 147 -1 N ILE A 146 O ILE A 110 LINK CA CA A 160 OD1 ASP A 103 LINK CA CA A 160 OD1 ASN A 105 LINK CA CA A 160 OD1 ASP A 107 LINK CA CA A 160 OE1 GLU A 114 LINK CA CA A 160 OE2 GLU A 114 LINK CA CA A 161 OD1 ASN A 141 LINK CA CA A 161 OE1 GLU A 150 CRYST1 46.900 46.900 152.300 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 0.021322 0.012310 0.000000 0.00000 SCALE2 0.000000 0.024620 0.000000 0.00000 SCALE3 0.000000 0.000000 0.006566 0.00000 (ATOM LINES ARE NOT SHOWN.) END