HEADER MUSCLE PROTEIN 14-MAY-97 1AJ4 TITLE STRUCTURE OF CALCIUM-SATURATED CARDIAC TROPONIN C, NMR, TITLE 2 1 STRUCTURE COMPND MOL_ID: 1; COMPND 2 MOLECULE: TROPONIN C; COMPND 3 CHAIN: NULL; COMPND 4 SYNONYM: CTNC; COMPND 5 ENGINEERED: YES; COMPND 6 MUTATION: C35S, C84S; COMPND 7 BIOLOGICAL_UNIT: MONOMER; COMPND 8 OTHER_DETAILS: CARDIAC TROPONIN C, CALCIUM IONS BOUND AT COMPND 9 SITES II, III AND IV SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: GALLUS GALLUS; SOURCE 3 ORGANISM_COMMON: CHICKEN; SOURCE 4 ORGAN: HEART; SOURCE 5 TISSUE: MUSCLE; SOURCE 6 CELLULAR_LOCATION: THIN FILAMENT; SOURCE 7 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 8 EXPRESSION_SYSTEM_STRAIN: BL21 (DE3) PLYSS; SOURCE 9 EXPRESSION_SYSTEM_PLASMID: PET-23D; SOURCE 10 EXPRESSION_SYSTEM_GENE: CTNC(A-CYS) KEYWDS CARDIAC, MUSCLE PROTEIN, REGULATORY, CALCIUM BINDING EXPDTA NMR, 1 STRUCTURE AUTHOR S.K.SIA,M.X.LI,L.SPYRACOPOULOS,S.M.GAGNE,W.LIU,J.A.PUTKEY, AUTHOR 2 B.D.SYKES REVDAT 1 20-MAY-98 1AJ4 0 JRNL AUTH S.K.SIA,M.X.LI,L.SPYRACOPOULOS,S.M.GAGNE,W.LIU, JRNL AUTH 2 J.A.PUTKEY,B.D.SYKES JRNL TITL STRUCTURE OF CARDIAC MUSCLE TROPONIN C UNEXPECTEDLY JRNL TITL 2 REVEALS A CLOSED REGULATORY DOMAIN JRNL REF J.BIOL.CHEM. V. 272 18216 1997 JRNL REFN ASTM JBCHA3 US ISSN 0021-9258 0071 REMARK 1 REMARK 2 REMARK 2 RESOLUTION. NOT APPLICABLE. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : X-PLOR 3.1 REMARK 3 AUTHORS : BRUNGER REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: REFINEMENT DETAILS CAN BE FOUND REMARK 3 IN THE JRNL CITATION. REMARK 3 REMARK 3 RMSD'S FOR BACKBONE ATOMS OF WELL-DEFINED REGIONS ARE: REMARK 3 0.54 (N-DOMAIN) AND 0.46 (C-DOMAIN). THE RMSD'S FOR ALL REMARK 3 HEAVY ATOMS OF WELL-DEFINED REGIONS: 0.97 (N-DOMAIN) AND REMARK 3 0.94 (C-DOMAIN). REMARK 4 REMARK 4 1AJ4 COMPLIES WITH FORMAT V. 2.2, 16-DEC-1996 REMARK 6 REMARK 6 CALCULATIONS OF THE N- AND C-DOMAINS WERE PERFORMED REMARK 6 SEPARATELY BECAUSE THE CENTRAL LINKER IS UNSTRUCTURED AND REMARK 6 FLEXIBLE IN SOLUTION. THE ORIENTATION OF THE TWO DOMAINS REMARK 6 WITH RESPECT TO EACH OTHER AND THE STRUCTURE OF THE REMARK 6 CENTRAL LINKER (RESIDUES 86 - 94) ARE THEREFORE UNDEFINED. REMARK 6 THE N-DOMAIN (2 - 85) AND C-DOMAIN (95 - 161) IN THIS REMARK 6 STRUCTURE ARE THE STRUCTURES IN THE ENSEMBLES CLOSEST TO REMARK 6 THE UNMINIMIZED AVERAGE STRUCTURES. IN PARTICULAR, THE REMARK 6 N-DOMAIN HERE CORRESPONDS TO MODEL 25 IN 2CTN, AND THE REMARK 6 C-DOMAIN TO MODEL 3 IN 3CTN. THE CENTRAL LINKER HERE IS REMARK 6 THE RESULT OF ENERGY MINIMIZATION WITHOUT EXPERIMENTAL REMARK 6 RESTRAINTS, AND IS ADDED ONLY AS A TOOL OF CONVENIENCE. REMARK 210 REMARK 210 EXPERIMENTAL DETAILS REMARK 210 EXPERIMENT TYPE : NMR REMARK 210 TEMPERATURE (KELVIN) : 303 REMARK 210 PH : 6.7 REMARK 210 REMARK 210 NMR EXPERIMENTS CONDUCTED : NOESY AND TOCSY REMARK 210 SPECTROMETER FIELD STRENGTH : 500 AND 600 REMARK 210 SPECTROMETER MODEL : UNITY PLUS REMARK 210 SPECTROMETER MANUFACTURER : VARIAN REMARK 210 REMARK 210 STRUCTURE DETERMINATION. REMARK 210 SOFTWARE USED : X-PLOR REMARK 210 METHOD USED : SIMULATED ANNEALING REMARK 210 REMARK 210 CONFORMERS, NUMBER CALCULATED : 35 REMARK 210 CONFORMERS, NUMBER SUBMITTED : 1 REMARK 210 CONFORMERS, SELECTION CRITERIA : SMALLEST RMSD TO THE REMARK 210 AVERAGE STRUCTURE REMARK 210 REMARK 210 REMARK: REMARK 210 IONIC_STRENGTH: NULL REMARK 210 PRESSURE: NULL REMARK 210 SOLVENT SYSTEM: H2O REMARK 210 REMARK 210 THE STRUCTURE WAS DETERMINED USING TRIPLE-RESONANCE NMR REMARK 210 SPECTROSCOPY ON 13C, 15N-LABELED INTACT CARDIAC TROPONIN C REMARK 210 IN THE CALCIUM-SATURATED STATE. REMARK 215 REMARK 215 NMR STUDY REMARK 215 THE COORDINATES IN THIS ENTRY WERE GENERATED FROM SOLUTION REMARK 215 NMR DATA. PROTEIN DATA BANK CONVENTIONS REQUIRE THAT REMARK 215 CRYST1 AND SCALE RECORDS BE INCLUDED, BUT THE VALUES ON REMARK 215 THESE RECORDS ARE MEANINGLESS. REMARK 600 REMARK 600 HETEROGEN REMARK 600 CA: CALCIUM IONS ARE BOUND AT SITES II, III, AND IV. REMARK 650 REMARK 650 HELIX REMARK 650 DETERMINATION METHOD: AUTHOR-DETERMINED REMARK 700 REMARK 700 SHEET REMARK 700 DETERMINATION METHOD: AUTHOR-DETERMINED REMARK 850 REMARK 850 CORRECTION BEFORE RELEASE REMARK 850 ORIGINAL DEPOSITION REVISED PRIOR TO RELEASE REMARK 850 DATE REVISED: 29-MAY-1997 TRACKING NUMBER: T11596 REMARK 999 REMARK 999 SEQUENCE REMARK 999 1AJ4 SWS P09860 1 - 1 NOT IN ATOMS LIST DBREF 1AJ4 2 161 SWS P09860 TPCC_CHICK 2 161 SEQADV 1AJ4 ALA 2 SWS P09860 ASP 2 CONFLICT SEQADV 1AJ4 SER 35 SWS P09860 CYS 35 ENGINEERED SEQADV 1AJ4 SER 84 SWS P09860 CYS 84 ENGINEERED SEQRES 1 161 ALA ALA ASP ILE TYR LYS ALA ALA VAL GLU GLN LEU THR SEQRES 2 161 GLU GLU GLN LYS ASN GLU PHE LYS ALA ALA PHE ASP ILE SEQRES 3 161 PHE VAL LEU GLY ALA GLU ASP GLY SER ILE SER THR LYS SEQRES 4 161 GLU LEU GLY LYS VAL MET ARG MET LEU GLY GLN ASN PRO SEQRES 5 161 THR PRO GLU GLU LEU GLN GLU MET ILE ASP GLU VAL ASP SEQRES 6 161 GLU ASP GLY SER GLY THR VAL ASP PHE ASP GLU PHE LEU SEQRES 7 161 VAL MET MET VAL ARG SER MET LYS ASP ASP SER LYS GLY SEQRES 8 161 LYS THR GLU GLU GLU LEU SER ASP LEU PHE ARG MET PHE SEQRES 9 161 ASP LYS ASN ALA ASP GLY TYR ILE ASP LEU GLU GLU LEU SEQRES 10 161 LYS ILE MET LEU GLN ALA THR GLY GLU THR ILE THR GLU SEQRES 11 161 ASP ASP ILE GLU GLU LEU MET LYS ASP GLY ASP LYS ASN SEQRES 12 161 ASN ASP GLY ARG ILE ASP TYR ASP GLU PHE LEU GLU PHE SEQRES 13 161 MET LYS GLY VAL GLU HET CA 1 1 HET CA 2 1 HET CA 3 1 HETNAM CA CALCIUM ION FORMUL 2 CA 3(CA1 2+) HELIX 1 HN LYS 6 GLU 10 1 5 HELIX 2 HA GLU 14 VAL 28 1 15 HELIX 3 HB THR 38 LEU 48 1 11 HELIX 4 HC PRO 54 VAL 64 1 11 HELIX 5 HD PHE 74 ARG 83 1 10 HELIX 6 HE GLU 95 MET 103 1 9 HELIX 7 HF LEU 114 ALA 123 1 10 HELIX 8 HG GLU 130 GLY 140 1 11 HELIX 9 HH TYR 150 LYS 158 1 9 SHEET 1 S1 2 SER 35 SER 37 0 SHEET 2 S1 2 THR 71 ASP 73 -1 N VAL 72 O ILE 36 SHEET 1 S2 2 TYR 111 ASP 113 0 SHEET 2 S2 2 ARG 147 ASP 149 -1 O ILE 148 N ILE 112 LINK CA CA 1 CG ASP 65 LINK CA CA 1 OD1 ASP 65 LINK CA CA 1 OD2 ASP 65 LINK CA CA 2 OD1 ASP 109 LINK CA CA 2 O TYR 111 LINK CA CA 3 O ARG 147 LINK CA CA 3 OE2 GLU 152 CRYST1 1.000 1.000 1.000 90.00 90.00 90.00 P 1 1 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 1.000000 0.000000 0.000000 0.00000 SCALE2 0.000000 1.000000 0.000000 0.00000 SCALE3 0.000000 0.000000 1.000000 0.00000 (ATOM LINES ARE NOT SHOWN.) END