HEADER CALMODULIN 18-MAR-99 1CFF TITLE NMR SOLUTION STRUCTURE OF A COMPLEX OF CALMODULIN WITH A TITLE 2 BINDING PEPTIDE OF THE CA2+-PUMP COMPND MOL_ID: 1; COMPND 2 MOLECULE: CALMODULIN; COMPND 3 CHAIN: A; COMPND 4 SYNONYM: CAM; COMPND 5 ENGINEERED: YES; COMPND 6 MOL_ID: 2; COMPND 7 MOLECULE: CALCIUM PUMP; COMPND 8 CHAIN: B; COMPND 9 FRAGMENT: CAM-BINDING DOMAIN; COMPND 10 SYNONYM: C20W; COMPND 11 EC: 3.6.1.38; COMPND 12 OTHER_DETAILS: SYNTHETIC SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: XENOPUS LAEVIS; SOURCE 3 ORGANISM_COMMON: AFRICAN CLAWED FROG; SOURCE 4 STRAIN: 71; SOURCE 5 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 6 EXPRESSION_SYSTEM_STRAIN: AR58; SOURCE 7 EXPRESSION_SYSTEM_CELLULAR_LOCATION: CYTOPLASM; SOURCE 8 EXPRESSION_SYSTEM_VECTOR: PTNCO12; SOURCE 9 EXPRESSION_SYSTEM_GENE: CALMODULIN; SOURCE 10 MOL_ID: 2; SOURCE 11 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 12 ORGANISM_COMMON: HUMAN; SOURCE 13 CELL: ERYTHROCYTE; SOURCE 14 CELLULAR_LOCATION: MEMBRANE; SOURCE 15 OTHER_DETAILS: SYNTHETIC KEYWDS CALMODULIN, C20W, PLASMA MEMBRANE CALCIUM PUMP, NMR EXPDTA NMR, 26 STRUCTURES AUTHOR B.ELSHORST,M.HENNIG,H.FOERSTERLING,A.DIENER,M.MAURER, AUTHOR 2 P.SCHULTE,H.SCHWALBE,J.KREBS,H.SCHMID,T.VORHERR,E.CARAFOLI, AUTHOR 3 C.GRIESINGER REVDAT 3 10-NOV-99 1CFF 1 SEQADV REMARK JRNL REVDAT 2 21-OCT-99 1CFF 1 REVDAT REVDAT 1 24-SEP-99 1CFF 0 JRNL AUTH B.ELSHORST,M.HENNIG,H.FOERSTERLING,A.DIENER, JRNL AUTH 2 M.MAURER,P.SCHULTE,H.SCHWALBE,C.GRIESINGER,J.KREBS, JRNL AUTH 3 H.SCHMID,T.VORHERR,E.CARAFOLI JRNL TITL NMR SOLUTION STRUCTURE OF A COMPLEX OF CALMODULIN JRNL TITL 2 WITH A BINDING PEPTIDE OF THE CA2+ PUMP JRNL REF BIOCHEMISTRY V. 38 12320 1999 JRNL REFN ASTM BICHAW US ISSN 0006-2960 REMARK 1 REMARK 2 REMARK 2 RESOLUTION. NOT APPLICABLE. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : X-PLOR 3.851 REMARK 3 AUTHORS : BRUNGER REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: REFINEMENT DETAILS CAN BE FOUND REMARK 3 IN THE JRNL CITATION ABOVE. IT IS IMPOSSIBLE TO REMARK 3 SUPERIMPOSE THE N- AND C-TERMINAL DOMAIN OF CAM REMARK 3 SIMULTANEOUSLY DUE TO THE LACK OF NOES BETWEEN BOTH REMARK 3 DOMAINS. THEREFOR THE 26 STRUCTURES ARE SUPERIMPOSED ONLY REMARK 3 OVER THE BACKBONE ATOMS OF THE C-TERMINAL DOMAIN OF CAM REMARK 3 AND THE PEPTIDE C20W. RMSD VALUES OF THE N-TERMINAL REMARK 3 DOMAIN OF CAM (LEU A 4 - ALA A 73) ARE 0.85 (BACKBONE) REMARK 3 AND 1.27 (HEAVY ATOMS) ANGSTROMS. RMSD VALUE FOR THE C- REMARK 3 TERMINAL DOMAIN OF CAM AND THE PEPTIDE C20W (ILE A 85 - REMARK 3 MET A 145, GLY B 4 - GLN B 16) ARE 0.57 (BACKBONE) AND REMARK 3 1.08 (HEAVY ATOMS) ANGSTROMS. RESIDUES ALA A 1 - GLN A 3, REMARK 3 ARG A 74 - GLU A 84, THR A 146 - LYS A 148 OF CAM AND REMARK 3 RESIDUES LEU B 1 - ARG B 3, THR B 17 - LYS B 20 OF THE REMARK 3 PEPTIDE C20W ARE DISORDERED. REMARK 4 REMARK 4 1CFF COMPLIES WITH FORMAT V. 2.3, 09-JULY-1998 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 23-MAR-1999. REMARK 100 THE RCSB ID CODE IS RCSB000682. REMARK 210 REMARK 210 EXPERIMENTAL DETAILS REMARK 210 EXPERIMENT TYPE : NMR REMARK 210 TEMPERATURE (KELVIN) : 303.00 REMARK 210 PH : 6.50 REMARK 210 IONIC STRENGTH : 0.115 REMARK 210 PRESSURE : 1 ATM REMARK 210 SAMPLE CONTENTS : 90% H2O/10% D2O, 100% D2O REMARK 210 REMARK 210 NMR EXPERIMENTS CONDUCTED : HNCO, HNCA, CBCACONH, REMARK 210 CBCANH, HCCCONH, HCCH- REMARK 210 TOCSY AND NOESY-HSQC REMARK 210 SPECTROMETER FIELD STRENGTH : 600 MHZ, 800 MHZ REMARK 210 SPECTROMETER MODEL : DRX600, DRX800 REMARK 210 SPECTROMETER MANUFACTURER : BRUKER REMARK 210 REMARK 210 STRUCTURE DETERMINATION. REMARK 210 SOFTWARE USED : NULL REMARK 210 METHOD USED : SIMULATED ANNEALING REMARK 210 REMARK 210 CONFORMERS, NUMBER CALCULATED : 200 REMARK 210 CONFORMERS, NUMBER SUBMITTED : 26 REMARK 210 CONFORMERS, SELECTION CRITERIA : LOWEST ENERGY REMARK 210 REMARK 210 BEST REPRESENTATIVE CONFORMER IN THIS ENSEMBLE : NULL REMARK 210 REMARK 210 REMARK: THE UNLABELED PEPTIDE C20W WAS ASSIGNED USING A REMARK 210 12C,14N-FILTERED NOESY- EXPERIMENT. INTERMOLECULAR NOES REMARK 210 BETWEEN 13C,15N-LABELED CALMODULIN AND THE PEPTIDE WERE REMARK 210 MEASURED USING A 12C,14N-F2-FILTERED NOESY-HSQC EXPERIMENT 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 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: COVALENT BOND ANGLES REMARK 500 REMARK 500 THE STEREOCHEMICAL PARAMETERS OF THE FOLLOWING RESIDUES REMARK 500 HAVE VALUES WHICH DEVIATE FROM EXPECTED VALUES BY MORE REMARK 500 THAN 6*RMSD (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN REMARK 500 IDENTIFIER; SSEQ=SEQUENCE NUMBER; I=INSERTION CODE). REMARK 500 REMARK 500 STANDARD TABLE: REMARK 500 FORMAT: (10X,I3,1X,A3,1X,A1,I4,A1,3(1X,A4,2X),12X,F5.1) REMARK 500 REMARK 500 EXPECTED VALUES: ENGH AND HUBER, 1991 REMARK 500 REMARK 500 M RES CSSEQI ATM1 ATM2 ATM3 REMARK 500 1 PHE B 9 N - CA - C ANGL. DEV. = -4.7 DEGREES REMARK 500 2 PHE B 9 N - CA - C ANGL. DEV. = -4.5 DEGREES REMARK 500 3 PHE B 9 N - CA - C ANGL. DEV. = -4.2 DEGREES REMARK 500 4 PHE B 9 N - CA - C ANGL. DEV. = -4.3 DEGREES REMARK 500 5 PHE B 9 N - CA - C ANGL. DEV. = -4.7 DEGREES REMARK 500 7 PHE B 9 N - CA - C ANGL. DEV. = -4.7 DEGREES REMARK 500 8 PHE B 9 N - CA - C ANGL. DEV. = -4.4 DEGREES REMARK 500 10 PHE B 9 N - CA - C ANGL. DEV. = -5.2 DEGREES REMARK 500 11 PHE B 9 N - CA - C ANGL. DEV. = -4.3 DEGREES REMARK 500 12 PHE B 9 N - CA - C ANGL. DEV. = -4.3 DEGREES REMARK 500 13 PHE B 9 N - CA - C ANGL. DEV. = -4.5 DEGREES REMARK 500 14 PHE B 9 N - CA - C ANGL. DEV. = -4.8 DEGREES REMARK 500 16 ILE A 9 N - CA - CB ANGL. DEV. = -4.3 DEGREES REMARK 500 16 PHE B 9 N - CA - C ANGL. DEV. = -4.2 DEGREES REMARK 500 18 PHE B 9 N - CA - C ANGL. DEV. = -4.5 DEGREES REMARK 500 20 PHE B 9 N - CA - C ANGL. DEV. = -4.3 DEGREES REMARK 500 21 THR A 34 CA - CB - CG2 ANGL. DEV. = 4.4 DEGREES REMARK 500 21 PHE B 9 N - CA - C ANGL. DEV. = -5.4 DEGREES REMARK 500 22 ILE A 9 N - CA - CB ANGL. DEV. = -4.7 DEGREES REMARK 500 22 PHE B 9 N - CA - C ANGL. DEV. = -5.0 DEGREES REMARK 500 23 PHE B 9 N - CA - C ANGL. DEV. = -4.7 DEGREES REMARK 500 24 THR A 34 CA - CB - CG2 ANGL. DEV. = 4.5 DEGREES REMARK 500 24 GLU A 87 N - CA - C ANGL. DEV. = -4.8 DEGREES REMARK 500 24 PHE B 9 N - CA - C ANGL. DEV. = -4.4 DEGREES REMARK 500 25 THR A 110 CA - CB - CG2 ANGL. DEV. = 5.4 DEGREES REMARK 500 25 PHE B 9 N - CA - C ANGL. DEV. = -4.4 DEGREES REMARK 500 26 PHE B 9 N - CA - C ANGL. DEV. = -4.8 DEGREES REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: TORSION ANGLES REMARK 500 REMARK 500 TORSION ANGLES OUTSIDE THE EXPECTED RAMACHANDRAN REGIONS: REMARK 500 (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN IDENTIFIER; REMARK 500 SSEQ=SEQUENCE NUMBER; I=INSERTION CODE). REMARK 500 STANDARD TABLE: REMARK 500 FORMAT:(10X,I3,1X,A3,1X,A1,I4,A1,4X,F7.2,3X,F7.2) REMARK 500 REMARK 500 M RES CSSEQI PSI PHI REMARK 500 REMARK 500 2 LEU A 4 58.76 150.61 REMARK 500 3 GLN A 41 1.68 68.02 REMARK 500 3 LYS A 94 38.13 -91.09 REMARK 500 4 LYS A 94 30.17 -89.03 REMARK 500 5 LEU A 4 86.94 136.89 REMARK 500 6 THR A 79 76.91 -61.46 REMARK 500 7 GLN A 3 74.60 144.44 REMARK 500 7 LYS A 77 68.42 128.91 REMARK 500 7 ASP A 80 62.39 127.02 REMARK 500 7 LYS A 94 34.76 -89.85 REMARK 500 8 ASP A 2 71.92 121.32 REMARK 500 8 MET A 76 63.02 160.78 REMARK 500 8 GLU A 83 64.02 -72.33 REMARK 500 8 LYS A 94 33.13 -89.77 REMARK 500 8 ASP A 118 143.37 -30.06 REMARK 500 8 ALA A 147 59.96 168.57 REMARK 500 9 GLU A 82 66.18 -70.10 REMARK 500 9 GLU A 83 71.52 -67.50 REMARK 500 10 LYS A 94 32.39 -89.21 REMARK 500 10 ARG B 2 62.55 140.13 REMARK 500 11 ARG B 2 73.98 -59.45 REMARK 500 12 GLU A 83 66.49 -70.85 REMARK 500 12 LYS A 94 30.13 -85.12 REMARK 500 13 LYS A 94 35.76 -90.28 REMARK 500 14 ASP A 2 69.59 -68.80 REMARK 500 14 LEU A 4 63.24 125.99 REMARK 500 14 LYS A 77 59.11 160.17 REMARK 500 14 GLU A 84 79.18 -57.44 REMARK 500 14 LYS A 94 30.08 -89.18 REMARK 500 15 LYS A 94 38.14 -86.88 REMARK 500 15 ILE B 19 73.63 143.10 REMARK 500 16 GLU A 83 58.23 -86.91 REMARK 500 16 ILE B 19 53.45 157.55 REMARK 500 17 ASP A 78 63.45 -74.68 REMARK 500 17 GLU A 82 77.59 -72.35 REMARK 500 17 GLU A 83 62.83 -79.67 REMARK 500 18 ASP A 2 63.09 124.79 REMARK 500 18 LYS A 94 34.81 -90.50 REMARK 500 19 LYS A 94 34.20 -94.03 REMARK 500 20 GLN A 3 61.08 145.51 REMARK 500 20 LEU A 4 62.16 143.03 REMARK 500 20 LYS A 94 38.89 -91.50 REMARK 500 20 ASP A 118 129.63 -29.16 REMARK 500 21 LYS A 94 31.72 -84.94 REMARK 500 22 ASP A 80 59.17 160.27 REMARK 500 22 GLU A 82 55.73 168.04 REMARK 500 23 MET A 76 70.41 158.49 REMARK 500 24 GLU A 82 60.67 -75.51 REMARK 500 24 GLU A 84 67.32 158.22 REMARK 500 24 LYS A 94 34.04 -90.16 REMARK 500 24 ALA A 147 58.55 162.63 REMARK 500 24 ARG B 2 60.83 -81.84 REMARK 500 24 ARG B 3 62.25 129.29 REMARK 500 25 THR A 5 55.86 138.46 REMARK 500 25 LYS A 94 36.05 -85.74 REMARK 500 26 GLU A 84 68.69 -60.62 REMARK 500 26 LYS A 94 31.06 -87.90 REMARK 650 REMARK 650 HELIX REMARK 650 DETERMINATION METHOD: AUTHOR-DETERMINED REMARK 700 REMARK 700 SHEET REMARK 700 DETERMINATION METHOD: AUTHOR-DETERMINED REMARK 800 REMARK 800 SITE REMARK 800 SITE_IDENTIFIER: CA1 REMARK 800 SITE_DESCRIPTION: REMARK 800 CALCIUM BINDING SITE REMARK 800 REMARK 800 SITE_IDENTIFIER: CA2 REMARK 800 SITE_DESCRIPTION: REMARK 800 CALCIUM BINDING SITE REMARK 800 REMARK 800 SITE_IDENTIFIER: CA3 REMARK 800 SITE_DESCRIPTION: REMARK 800 CALCIUM BINDING SITE REMARK 800 REMARK 800 SITE_IDENTIFIER: CA4 REMARK 800 SITE_DESCRIPTION: REMARK 800 CALCIUM BINDING SITE REMARK 800 DBREF 1CFF A 1 148 GB 214019 K01945 2 149 DBREF 1CFF B 1 20 GB 190133 J04027 1100 1119 SEQADV 1CFF LYS B 20 GB 190133 ARG 1119 MUTATION SEQRES 1 A 148 ALA ASP GLN LEU THR GLU GLU GLN ILE ALA GLU PHE LYS SEQRES 2 A 148 GLU ALA PHE SER LEU PHE ASP LYS ASP GLY ASP GLY THR SEQRES 3 A 148 ILE THR THR LYS GLU LEU GLY THR VAL MET ARG SER LEU SEQRES 4 A 148 GLY GLN ASN PRO THR GLU ALA GLU LEU GLN ASP MET ILE SEQRES 5 A 148 ASN GLU VAL ASP ALA ASP GLY ASN GLY THR ILE ASP PHE SEQRES 6 A 148 PRO GLU PHE LEU THR MET MET ALA ARG LYS MET LYS ASP SEQRES 7 A 148 THR ASP SER GLU GLU GLU ILE ARG GLU ALA PHE ARG VAL SEQRES 8 A 148 PHE ASP LYS ASP GLY ASN GLY TYR ILE SER ALA ALA GLU SEQRES 9 A 148 LEU ARG HIS VAL MET THR ASN LEU GLY GLU LYS LEU THR SEQRES 10 A 148 ASP GLU GLU VAL ASP GLU MET ILE ARG GLU ALA ASP ILE SEQRES 11 A 148 ASP GLY ASP GLY GLN VAL ASN TYR GLU GLU PHE VAL GLN SEQRES 12 A 148 MET MET THR ALA LYS SEQRES 1 B 20 LEU ARG ARG GLY GLN ILE LEU TRP PHE ARG GLY LEU ASN SEQRES 2 B 20 ARG ILE GLN THR GLN ILE LYS HET CA A 149 1 HET CA A 150 1 HET CA A 151 1 HET CA A 152 1 HETNAM CA CALCIUM ION FORMUL 3 CA 4(CA1 2+) HELIX 1 A GLU A 7 LEU A 18 1 12 HELIX 2 B THR A 29 SER A 38 1 10 HELIX 3 C GLU A 47 GLU A 54 1 8 HELIX 4 D PHE A 65 ALA A 73 1 9 HELIX 5 E ILE A 85 PHE A 92 1 8 HELIX 6 F ALA A 103 LEU A 112 1 10 HELIX 7 G ASP A 118 ALA A 128 1 11 HELIX 8 H TYR A 138 MET A 145 1 8 SHEET 1 S1 1 THR A 26 THR A 28 0 SHEET 1 S2 1 THR A 62 ASP A 64 0 SHEET 1 S3 1 TYR A 99 SER A 101 0 SHEET 1 S4 1 GLN A 135 ASN A 137 0 SITE 1 CA1 5 ASP A 20 ASP A 22 ASP A 24 THR A 26 SITE 2 CA1 5 GLU A 31 SITE 1 CA2 5 ASP A 56 ASP A 58 ASN A 60 THR A 62 SITE 2 CA2 5 GLU A 67 SITE 1 CA3 5 ASP A 93 ASP A 95 ASN A 97 TYR A 99 SITE 2 CA3 5 GLU A 104 SITE 1 CA4 5 ASP A 129 ASP A 131 ASP A 133 GLN A 135 SITE 2 CA4 5 GLU A 140 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 MODEL 1 (ATOM LINES ARE NOT SHOWN.) END