HEADER STRUCTURAL PROTEIN 16-DEC-09 2KRD
TITLE SOLUTION STRUCTURE OF THE REGULATORY DOMAIN OF HUMAN CARDIAC TROPONIN
TITLE 2 C IN COMPLEX WITH THE SWITCH REGION OF CARDIAC TROPONIN I AND W7
COMPND MOL_ID: 1;
COMPND 2 MOLECULE: TROPONIN C, SLOW SKELETAL AND CARDIAC MUSCLES;
COMPND 3 CHAIN: C;
COMPND 4 SYNONYM: TN-C;
COMPND 5 ENGINEERED: YES;
COMPND 6 MOL_ID: 2;
COMPND 7 MOLECULE: TROPONIN I, CARDIAC MUSCLE;
COMPND 8 CHAIN: I;
COMPND 9 SYNONYM: CARDIAC TROPONIN I;
COMPND 10 ENGINEERED: YES
SOURCE MOL_ID: 1;
SOURCE 2 ORGANISM_SCIENTIFIC: HOMO SAPIENS;
SOURCE 3 ORGANISM_COMMON: HUMAN;
SOURCE 4 ORGANISM_TAXID: 9606;
SOURCE 5 GENE: TNNC1, TNNC;
SOURCE 6 EXPRESSION_SYSTEM: ESCHERICHIA COLI;
SOURCE 7 EXPRESSION_SYSTEM_TAXID: 562;
SOURCE 8 EXPRESSION_SYSTEM_VECTOR_TYPE: PLASMID;
SOURCE 9 EXPRESSION_SYSTEM_PLASMID: PET3A;
SOURCE 10 MOL_ID: 2;
SOURCE 11 SYNTHETIC: YES;
SOURCE 12 ORGANISM_SCIENTIFIC: HOMO SAPIENS;
SOURCE 13 ORGANISM_COMMON: HUMAN;
SOURCE 14 ORGANISM_TAXID: 9606
KEYWDS CARDIAC TROPONIN C, REGULATORY DOMAIN, TROPONIN I, SWITCH REGION, W7,
KEYWDS 2 ACETYLATION, CALCIUM, CARDIOMYOPATHY, DISEASE MUTATION, MUSCLE
KEYWDS 3 PROTEIN, POLYMORPHISM, ACTIN-BINDING, PHOSPHOPROTEIN, STRUCTURAL
KEYWDS 4 PROTEIN
EXPDTA SOLUTION NMR
NUMMDL 20
AUTHOR M.OLESZCZUK,I.M.ROBERTSON,M.X.LI,B.D.SYKES
REVDAT 3 16-MAR-22 2KRD 1 REMARK LINK
REVDAT 2 28-APR-10 2KRD 1 JRNL
REVDAT 1 16-FEB-10 2KRD 0
JRNL AUTH M.OLESZCZUK,I.M.ROBERTSON,M.X.LI,B.D.SYKES
JRNL TITL SOLUTION STRUCTURE OF THE REGULATORY DOMAIN OF HUMAN CARDIAC
JRNL TITL 2 TROPONIN C IN COMPLEX WITH THE SWITCH REGION OF CARDIAC
JRNL TITL 3 TROPONIN I AND W7: THE BASIS OF W7 AS AN INHIBITOR OF
JRNL TITL 4 CARDIAC MUSCLE CONTRACTION.
JRNL REF J.MOL.CELL.CARDIOL. V. 48 925 2010
JRNL REFN ISSN 0022-2828
JRNL PMID 20116385
JRNL DOI 10.1016/J.YJMCC.2010.01.016
REMARK 2
REMARK 2 RESOLUTION. NOT APPLICABLE.
REMARK 3
REMARK 3 REFINEMENT.
REMARK 3 PROGRAM : VNMRJ, X-PLOR NIH
REMARK 3 AUTHORS : VARIAN (VNMRJ), SCHWIETERS, C. ET AL. (X-PLOR NIH)
REMARK 3
REMARK 3 OTHER REFINEMENT REMARKS: PROTONATED FORM OF W7 WAS USED DURING
REMARK 3 WATER REFINEMENT. POSITIVE CHARGE ON W7 TAIL (-CH2-NH3+ GROUP)
REMARK 3 WAS ADDED MANUALLY IN ANALOGY TO POSITIVE CHARGE ON -CH2-NH3+
REMARK 3 GROUP IN LYS+.
REMARK 4
REMARK 4 2KRD COMPLIES WITH FORMAT V. 3.30, 13-JUL-11
REMARK 100
REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 22-DEC-09.
REMARK 100 THE DEPOSITION ID IS D_1000101478.
REMARK 210
REMARK 210 EXPERIMENTAL DETAILS
REMARK 210 EXPERIMENT TYPE : NMR
REMARK 210 TEMPERATURE (KELVIN) : 303
REMARK 210 PH : 6.8
REMARK 210 IONIC STRENGTH : NULL
REMARK 210 PRESSURE : AMBIENT
REMARK 210 SAMPLE CONTENTS : 0.5 MM [U-13C; U-15N] CNTNC, 10
REMARK 210 MM IMIDAZOLE, 100 MM POTASSIUM CHLORIDE, 3 MM N-(6-AMINOHEXYL)-5-
REMARK 210 CHLORO-1-NAPHTHALENESULFONAMIDE, 2.1 MM CTNI(147-163), 15 MM
REMARK 210 DITHIOTHREITOL (DTT), 0.5 MM DSS, 10 MM CALCIUM CHLORIDE, 90%
REMARK 210 H2O/10% D2O; 0.8 MM [U-13C; U-15N] CNTNC, 10 MM IMIDAZOLE, 100
REMARK 210 MM POTASSIUM CHLORIDE, 9 MM CALCIUM CHLORIDE, 2.5 MM N-(6-
REMARK 210 AMINOHEXYL)-5-CHLORO-1-NAPHTHALENESULFONAMIDE, 4.2 MM CTNI(147-
REMARK 210 163), 15 MM DITHIOTHREITOL (DTT), 0.5 MM DSS, 90% H2O/10% D2O;
REMARK 210 0.5 MM [U-13C; U-15N] CNTNC, 10 MM IMIDAZOLE, 100 MM POTASSIUM
REMARK 210 CHLORIDE, 9 MM CALCIUM CHLORIDE, 2.8 MM N-(6-AMINOHEXYL)-5-
REMARK 210 CHLORO-1-NAPHTHALENESULFONAMIDE, 2 MM CTNI(147-163), 15 MM
REMARK 210 DITHIOTHREITOL (DTT), 0.2 MM DSS, 100% D2O; 0.7 MM [U-13C; U-15N]
REMARK 210 CNTNC, 10 MM IMIDAZOLE, 100 MM POTASSIUM CHLORIDE, 9 MM CALCIUM
REMARK 210 CHLORIDE, 2.9 MM N-(6-AMINOHEXYL)-5-CHLORO-1-
REMARK 210 NAPHTHALENESULFONAMIDE, 3.6 MM CTNI(147-163), 15 MM
REMARK 210 DITHIOTHREITOL (DTT), 0.4 MM DSS, 100% D2O; 0.5 MM [U-13C; U-15N]
REMARK 210 CNTNC, 10 MM IMIDAZOLE, 100 MM POTASSIUM CHLORIDE, 9 MM CALCIUM
REMARK 210 CHLORIDE, 3 MM N-(6-AMINOHEXYL)-5-CHLORO-1-
REMARK 210 NAPHTHALENESULFONAMIDE, 2.2 MM CTNI(147-163), 15 MM
REMARK 210 DITHIOTHREITOL (DTT), 0.2 MM DSS, 100% D2O
REMARK 210
REMARK 210 NMR EXPERIMENTS CONDUCTED : 2D 1H-15N HSQC; 2D 1H-13C HSQC;
REMARK 210 2D DQF-COSY; 3D CBCA(CO)NH; 3D
REMARK 210 C(CO)NH; 3D HNCACB; 3D 1H-15N
REMARK 210 NOESY; 3D 1H-13C NOESY; 2D 1H-1H
REMARK 210 NOESY; 2D CNFILNOESY; 2D
REMARK 210 CNFILTOCSY; 3D GCHMQCNOESY_
REMARK 210 CNFILT; 2D HBCBCGCDHDA; 3D HC(CO)
REMARK 210 HN
REMARK 210 SPECTROMETER FIELD STRENGTH : 800 MHZ; 600 MHZ; 500 MHZ
REMARK 210 SPECTROMETER MODEL : INOVA; UNITY
REMARK 210 SPECTROMETER MANUFACTURER : VARIAN
REMARK 210
REMARK 210 STRUCTURE DETERMINATION.
REMARK 210 SOFTWARE USED : NMRPIPE, NMRVIEW, CYANA, TALOS,
REMARK 210 X-PLOR NIH
REMARK 210 METHOD USED : SIMULATED ANNEALING
REMARK 210
REMARK 210 CONFORMERS, NUMBER CALCULATED : 300
REMARK 210 CONFORMERS, NUMBER SUBMITTED : 20
REMARK 210 CONFORMERS, SELECTION CRITERIA : STRUCTURES WITH THE LOWEST
REMARK 210 ENERGY
REMARK 210
REMARK 210 BEST REPRESENTATIVE CONFORMER IN THIS ENSEMBLE : 1
REMARK 210
REMARK 210 REMARK: NULL
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 300
REMARK 300 BIOMOLECULE: 1
REMARK 300 SEE REMARK 350 FOR THE AUTHOR PROVIDED AND/OR PROGRAM
REMARK 300 GENERATED ASSEMBLY INFORMATION FOR THE STRUCTURE IN
REMARK 300 THIS ENTRY. THE REMARK MAY ALSO PROVIDE INFORMATION ON
REMARK 300 BURIED SURFACE AREA.
REMARK 350
REMARK 350 COORDINATES FOR A COMPLETE MULTIMER REPRESENTING THE KNOWN
REMARK 350 BIOLOGICALLY SIGNIFICANT OLIGOMERIZATION STATE OF THE
REMARK 350 MOLECULE CAN BE GENERATED BY APPLYING BIOMT TRANSFORMATIONS
REMARK 350 GIVEN BELOW. BOTH NON-CRYSTALLOGRAPHIC AND
REMARK 350 CRYSTALLOGRAPHIC OPERATIONS ARE GIVEN.
REMARK 350
REMARK 350 BIOMOLECULE: 1
REMARK 350 AUTHOR DETERMINED BIOLOGICAL UNIT: DIMERIC
REMARK 350 APPLY THE FOLLOWING TO CHAINS: C, I
REMARK 350 BIOMT1 1 1.000000 0.000000 0.000000 0.00000
REMARK 350 BIOMT2 1 0.000000 1.000000 0.000000 0.00000
REMARK 350 BIOMT3 1 0.000000 0.000000 1.000000 0.00000
REMARK 500
REMARK 500 GEOMETRY AND STEREOCHEMISTRY
REMARK 500 SUBTOPIC: CLOSE CONTACTS
REMARK 500
REMARK 500 THE FOLLOWING ATOMS ARE IN CLOSE CONTACT.
REMARK 500
REMARK 500 ATM1 RES C SSEQI ATM2 RES C SSEQI DISTANCE
REMARK 500 O LEU C 41 H VAL C 44 1.54
REMARK 500 O SER C 37 H LYS C 39 1.56
REMARK 500
REMARK 500 REMARK: NULL
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
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 EXPECTED VALUES: GJ KLEYWEGT AND TA JONES (1996). PHI/PSI-
REMARK 500 CHOLOGY: RAMACHANDRAN REVISITED. STRUCTURE 4, 1395 - 1400
REMARK 500
REMARK 500 M RES CSSEQI PSI PHI
REMARK 500 1 VAL C 28 53.41 -103.53
REMARK 500 1 THR C 38 -38.19 61.46
REMARK 500 1 ASP C 87 79.56 19.43
REMARK 500 1 ASP C 88 84.42 13.88
REMARK 500 1 SER I 149 -142.61 -135.08
REMARK 500 1 ASP I 151 69.19 82.69
REMARK 500 1 LEU I 158 -64.43 53.45
REMARK 500 1 ALA I 162 -38.13 96.44
REMARK 500 2 VAL C 28 68.75 -112.95
REMARK 500 2 ASP C 33 -84.62 -87.96
REMARK 500 2 THR C 38 -39.20 65.59
REMARK 500 2 ASP C 65 82.95 -66.90
REMARK 500 2 ILE I 148 107.42 -52.05
REMARK 500 2 SER I 149 -56.40 67.05
REMARK 500 3 ASP C 2 -2.08 68.33
REMARK 500 3 CYS C 35 -161.33 56.21
REMARK 500 3 LYS C 86 -3.15 -58.19
REMARK 500 3 ASP C 87 121.10 -18.26
REMARK 500 3 ASP C 88 -153.49 28.62
REMARK 500 3 SER I 149 -52.70 60.82
REMARK 500 3 ALA I 156 -17.68 -47.22
REMARK 500 3 LEU I 158 -55.15 62.20
REMARK 500 3 ARG I 161 136.14 64.10
REMARK 500 3 ALA I 162 -104.24 67.10
REMARK 500 4 LEU C 29 46.01 36.20
REMARK 500 4 THR C 38 -33.00 66.59
REMARK 500 4 ASP C 88 -58.03 -142.94
REMARK 500 4 ILE I 148 -59.72 72.53
REMARK 500 4 SER I 149 -23.32 -159.78
REMARK 500 4 ASP I 151 44.97 -97.91
REMARK 500 4 LEU I 158 -86.30 25.33
REMARK 500 4 ALA I 162 -57.86 -120.35
REMARK 500 5 THR C 38 -39.73 61.51
REMARK 500 5 ASP C 87 134.15 79.74
REMARK 500 5 ILE I 148 -122.05 40.93
REMARK 500 5 ALA I 162 -80.64 179.21
REMARK 500 6 LEU C 29 70.45 23.45
REMARK 500 6 ASP C 87 -70.35 65.11
REMARK 500 6 ILE I 148 -79.70 38.05
REMARK 500 6 ALA I 152 -18.99 -47.01
REMARK 500 6 ALA I 160 78.02 -69.80
REMARK 500 7 CYS C 35 -172.00 58.32
REMARK 500 7 THR C 38 -33.66 85.33
REMARK 500 7 ASP C 88 -179.99 53.82
REMARK 500 7 SER I 149 -51.09 63.82
REMARK 500 7 ASP I 151 39.78 -93.63
REMARK 500 7 LEU I 158 -54.43 77.92
REMARK 500 7 ALA I 160 -10.68 69.70
REMARK 500 7 ALA I 162 -146.56 -80.46
REMARK 500 8 CYS C 35 -161.26 52.91
REMARK 500
REMARK 500 THIS ENTRY HAS 156 RAMACHANDRAN OUTLIERS.
REMARK 500
REMARK 500 REMARK: NULL
REMARK 620
REMARK 620 METAL COORDINATION
REMARK 620 (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN IDENTIFIER;
REMARK 620 SSEQ=SEQUENCE NUMBER; I=INSERTION CODE):
REMARK 620
REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL
REMARK 620 CA C 90 CA
REMARK 620 N RES CSSEQI ATOM
REMARK 620 1 ASP C 65 OD1
REMARK 620 2 ASP C 67 OD1 78.1
REMARK 620 3 SER C 69 OG 120.9 137.8
REMARK 620 4 THR C 71 O 73.7 148.1 71.6
REMARK 620 5 GLU C 76 OE1 94.9 90.5 121.4 77.5
REMARK 620 6 GLU C 76 OE2 139.2 81.0 97.9 111.4 50.5
REMARK 620 N 1 2 3 4 5
REMARK 800
REMARK 800 SITE
REMARK 800 SITE_IDENTIFIER: AC1
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE CA C 90
REMARK 800
REMARK 800 SITE_IDENTIFIER: AC2
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE WW7 I 91
DBREF 2KRD C 1 89 UNP P63316 TNNC1_HUMAN 1 89
DBREF 2KRD I 147 163 UNP P19429 TNNI3_HUMAN 148 164
SEQRES 1 C 89 MET ASP ASP ILE TYR LYS ALA ALA VAL GLU GLN LEU THR
SEQRES 2 C 89 GLU GLU GLN LYS ASN GLU PHE LYS ALA ALA PHE ASP ILE
SEQRES 3 C 89 PHE VAL LEU GLY ALA GLU ASP GLY CYS ILE SER THR LYS
SEQRES 4 C 89 GLU LEU GLY LYS VAL MET ARG MET LEU GLY GLN ASN PRO
SEQRES 5 C 89 THR PRO GLU GLU LEU GLN GLU MET ILE ASP GLU VAL ASP
SEQRES 6 C 89 GLU ASP GLY SER GLY THR VAL ASP PHE ASP GLU PHE LEU
SEQRES 7 C 89 VAL MET MET VAL ARG CYS MET LYS ASP ASP SER
SEQRES 1 I 17 ARG ILE SER ALA ASP ALA MET MET GLN ALA LEU LEU GLY
SEQRES 2 I 17 ALA ARG ALA LYS
HET CA C 90 1
HET WW7 I 91 44
HETNAM CA CALCIUM ION
HETNAM WW7 N-(6-AMINOHEXYL)-5-CHLORO-1-NAPHTHALENESULFONAMIDE
FORMUL 3 CA CA 2+
FORMUL 4 WW7 C16 H21 CL N2 O2 S
HELIX 1 1 ASP C 2 GLU C 10 1 9
HELIX 2 2 THR C 13 VAL C 28 1 16
HELIX 3 3 LEU C 41 GLY C 49 1 9
HELIX 4 4 THR C 53 ASP C 65 1 13
HELIX 5 5 PHE C 74 ASP C 87 1 14
HELIX 6 6 ASP I 151 ALA I 156 1 6
SHEET 1 A 2 CYS C 35 ILE C 36 0
SHEET 2 A 2 VAL C 72 ASP C 73 -1 O VAL C 72 N ILE C 36
LINK OD1 ASP C 65 CA CA C 90 1555 1555 1.98
LINK OD1 ASP C 67 CA CA C 90 1555 1555 2.33
LINK OG SER C 69 CA CA C 90 1555 1555 2.10
LINK O THR C 71 CA CA C 90 1555 1555 2.42
LINK OE1 GLU C 76 CA CA C 90 1555 1555 1.99
LINK OE2 GLU C 76 CA CA C 90 1555 1555 2.76
SITE 1 AC1 5 ASP C 65 ASP C 67 SER C 69 THR C 71
SITE 2 AC1 5 GLU C 76
SITE 1 AC2 7 MET C 47 MET C 60 VAL C 64 PHE C 77
SITE 2 AC2 7 ILE I 148 SER I 149 MET I 153
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
