HEADER SIGNALING PROTEIN 14-AUG-15 2N6A
TITLE NMR STRUCTURE OF A HUMAN CALMODULIN/CONNEXIN-36 PEPTIDE HYBRID
COMPND MOL_ID: 1;
COMPND 2 MOLECULE: HUMAN CALMODULIN/CONNEXIN-36 PEPTIDE HYBRID;
COMPND 3 CHAIN: A;
COMPND 4 SYNONYM: CAM, GJD2, CONNEXIN-36, CX36, GAP JUNCTION ALPHA-9 PROTEIN;
COMPND 5 ENGINEERED: YES
SOURCE MOL_ID: 1;
SOURCE 2 ORGANISM_SCIENTIFIC: HOMO SAPIENS;
SOURCE 3 ORGANISM_COMMON: HUMAN;
SOURCE 4 ORGANISM_TAXID: 9606;
SOURCE 5 EXPRESSION_SYSTEM: ESCHERICHIA COLI;
SOURCE 6 EXPRESSION_SYSTEM_TAXID: 562;
SOURCE 7 EXPRESSION_SYSTEM_VECTOR_TYPE: PLASMID;
SOURCE 8 EXPRESSION_SYSTEM_PLASMID: PD441
KEYWDS CALMODULIN, CALCIUM BINDING, GAP JUNCTION, CHIMERA, SIGNALING PROTEIN
EXPDTA SOLUTION NMR
NUMMDL 18
AUTHOR L.DONALDSON
REVDAT 3 14-JUN-23 2N6A 1 REMARK SEQADV LINK
REVDAT 2 27-APR-16 2N6A 1 DBREF REMARK SEQRES
REVDAT 1 26-AUG-15 2N6A 0
JRNL AUTH L.DONALDSON
JRNL TITL AN INVESTIGATION OF CALCIUM MEDIATE SIGNALING BY THE
JRNL TITL 2 CONNEXIN 36 GAP JUNCTION PROTEIN
JRNL REF TO BE PUBLISHED
JRNL REFN
REMARK 2
REMARK 2 RESOLUTION. NOT APPLICABLE.
REMARK 3
REMARK 3 REFINEMENT.
REMARK 3 PROGRAM : NMRPIPE, CYANA
REMARK 3 AUTHORS : DELAGLIO, GRZESIEK, VUISTER, ZHU, PFEIFER AND BAX
REMARK 3 (NMRPIPE), GUNTERT, MUMENTHALER AND WUTHRICH
REMARK 3 (CYANA)
REMARK 3
REMARK 3 OTHER REFINEMENT REMARKS: 10000 STEPS OF SA USING CYANA3
REMARK 4
REMARK 4 2N6A COMPLIES WITH FORMAT V. 3.30, 13-JUL-11
REMARK 100
REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 18-AUG-15.
REMARK 100 THE DEPOSITION ID IS D_1000104475.
REMARK 210
REMARK 210 EXPERIMENTAL DETAILS
REMARK 210 EXPERIMENT TYPE : NMR
REMARK 210 TEMPERATURE (KELVIN) : 293
REMARK 210 PH : 7.4
REMARK 210 IONIC STRENGTH : 0.25
REMARK 210 PRESSURE : AMBIENT
REMARK 210 SAMPLE CONTENTS : 2.0 MM [U-99% 13C; U-99% 15N]
REMARK 210 PROTEIN, 0.0005 W/V SODIUM AZIDE,
REMARK 210 25 MM SODIUM CHLORIDE, 5 MM
REMARK 210 TRIS, 90% H2O/10% D2O
REMARK 210
REMARK 210 NMR EXPERIMENTS CONDUCTED : 2D 1H-15N HSQC; 2D 1H-13C HSQC
REMARK 210 ALIPHATIC; 2D 1H-13C HSQC
REMARK 210 AROMATIC; 3D CBCA(CO)NH; 3D C(CO)
REMARK 210 NH; 3D HNCO; 3D HNCA; 3D HNCACB;
REMARK 210 3D H(CCO)NH; 3D HCCH-TOCSY; 3D
REMARK 210 1H-15N NOESY; 3D 1H-13C NOESY
REMARK 210 ALIPHATIC; 3D 1H-13C NOESY
REMARK 210 AROMATIC
REMARK 210 SPECTROMETER FIELD STRENGTH : 700 MHZ
REMARK 210 SPECTROMETER MODEL : AVANCE
REMARK 210 SPECTROMETER MANUFACTURER : BRUKER
REMARK 210
REMARK 210 STRUCTURE DETERMINATION.
REMARK 210 SOFTWARE USED : CYANA
REMARK 210 METHOD USED : SIMULATED ANNEALING
REMARK 210
REMARK 210 CONFORMERS, NUMBER CALCULATED : 400
REMARK 210 CONFORMERS, NUMBER SUBMITTED : 18
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: MONOMERIC
REMARK 350 APPLY THE FOLLOWING TO CHAINS: A
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 465
REMARK 465 MISSING RESIDUES
REMARK 465 THE FOLLOWING RESIDUES WERE NOT LOCATED IN THE
REMARK 465 EXPERIMENT. (RES=RESIDUE NAME; C=CHAIN IDENTIFIER;
REMARK 465 SSSEQ=SEQUENCE NUMBER; I=INSERTION CODE.)
REMARK 465 MODELS 1-18
REMARK 465 RES C SSSEQI
REMARK 465 HIS A -1
REMARK 465 HIS A 0
REMARK 465 HIS A 1
REMARK 465 HIS A 2
REMARK 465 HIS A 3
REMARK 465 HIS A 4
REMARK 465 THR A 5
REMARK 465 GLU A 6
REMARK 465 GLU A 7
REMARK 465 ARG A 170
REMARK 465 LYS A 171
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 ASN A 42 71.53 -119.07
REMARK 500 1 ILE A 85 -70.98 -51.35
REMARK 500 1 GLU A 120 -70.13 -51.80
REMARK 500 1 ALA A 152 33.05 -87.30
REMARK 500 2 LEU A 18 -70.67 -49.56
REMARK 500 2 ASP A 20 93.50 -62.95
REMARK 500 2 ASN A 42 72.16 -117.00
REMARK 500 2 ASP A 80 103.25 -51.81
REMARK 500 2 ILE A 85 -70.58 -49.89
REMARK 500 2 THR A 146 50.15 -113.38
REMARK 500 2 ALA A 152 32.00 -88.67
REMARK 500 3 ASN A 42 73.67 -113.74
REMARK 500 3 ILE A 85 -71.58 -52.41
REMARK 500 3 ALA A 152 33.22 -89.46
REMARK 500 4 ASN A 42 77.24 -116.55
REMARK 500 4 ASP A 80 109.63 -58.06
REMARK 500 4 ILE A 85 -70.54 -51.72
REMARK 500 4 GLU A 114 96.41 -61.01
REMARK 500 4 ALA A 152 33.44 -87.11
REMARK 500 5 ASP A 78 -178.63 -58.57
REMARK 500 5 THR A 79 -69.82 -97.50
REMARK 500 5 ILE A 85 -71.32 -49.36
REMARK 500 5 GLU A 120 -70.46 -58.31
REMARK 500 5 ALA A 152 33.15 -88.78
REMARK 500 6 ILE A 85 -70.57 -53.63
REMARK 500 6 ALA A 152 33.55 -87.54
REMARK 500 7 ASN A 42 70.16 -119.81
REMARK 500 7 SER A 81 -70.50 -97.06
REMARK 500 7 ILE A 85 -71.46 -51.61
REMARK 500 7 ALA A 152 32.35 -89.51
REMARK 500 8 PHE A 19 -70.40 -81.24
REMARK 500 8 ASP A 20 103.48 -59.50
REMARK 500 8 GLU A 31 -70.39 -60.46
REMARK 500 8 ILE A 85 -70.76 -49.57
REMARK 500 8 ALA A 103 -71.27 -66.28
REMARK 500 8 ALA A 152 34.10 -86.12
REMARK 500 9 ASN A 42 68.48 -117.70
REMARK 500 9 ILE A 85 -70.73 -48.71
REMARK 500 9 ALA A 151 -176.83 -66.43
REMARK 500 9 ALA A 152 34.45 -84.74
REMARK 500 9 SER A 154 -70.09 -52.41
REMARK 500 10 ASN A 42 72.44 -114.22
REMARK 500 10 ILE A 85 -70.96 -48.79
REMARK 500 10 ALA A 152 33.57 -87.23
REMARK 500 10 ALA A 168 -69.55 -93.61
REMARK 500 11 ASN A 42 72.97 -113.87
REMARK 500 11 ILE A 85 -71.04 -49.36
REMARK 500 11 ALA A 103 -70.79 -64.12
REMARK 500 11 GLU A 120 -70.07 -51.84
REMARK 500 11 ALA A 152 32.08 -90.98
REMARK 500
REMARK 500 THIS ENTRY HAS 89 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 A 201 CA
REMARK 620 N RES CSSEQI ATOM
REMARK 620 1 ASP A 20 OD1
REMARK 620 2 ASP A 22 OD2 69.5
REMARK 620 3 ASP A 24 OD2 81.5 69.9
REMARK 620 4 THR A 26 O 64.7 131.0 86.7
REMARK 620 5 GLU A 31 OE1 54.4 97.7 135.2 69.1
REMARK 620 6 GLU A 31 OE2 50.1 56.3 115.8 102.6 42.0
REMARK 620 N 1 2 3 4 5
REMARK 620
REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL
REMARK 620 CA A 202 CA
REMARK 620 N RES CSSEQI ATOM
REMARK 620 1 ASP A 56 OD1
REMARK 620 2 ASP A 58 OD1 63.2
REMARK 620 3 ASN A 60 OD1 54.5 69.8
REMARK 620 4 THR A 62 O 56.7 119.4 70.0
REMARK 620 5 GLU A 67 OE2 61.0 69.6 113.7 87.9
REMARK 620 6 GLU A 67 OE1 69.2 109.3 117.3 56.4 41.9
REMARK 620 N 1 2 3 4 5
REMARK 620
REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL
REMARK 620 CA A 203 CA
REMARK 620 N RES CSSEQI ATOM
REMARK 620 1 ASP A 93 OD1
REMARK 620 2 ASP A 95 OD2 70.5
REMARK 620 3 ASN A 97 OD1 49.8 68.3
REMARK 620 4 TYR A 99 O 56.7 126.0 69.6
REMARK 620 5 GLU A 104 OE1 65.3 97.5 114.9 71.6
REMARK 620 6 GLU A 104 OE2 50.1 55.9 91.6 93.6 42.0
REMARK 620 N 1 2 3 4 5
REMARK 620
REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL
REMARK 620 CA A 204 CA
REMARK 620 N RES CSSEQI ATOM
REMARK 620 1 ASP A 129 OD1
REMARK 620 2 ASP A 131 OD1 69.2
REMARK 620 3 ASP A 133 OD2 66.1 69.8
REMARK 620 4 GLN A 135 O 64.4 133.5 88.7
REMARK 620 5 GLU A 140 OE2 69.3 103.5 134.2 62.7
REMARK 620 6 GLU A 140 OE1 80.4 70.0 134.5 104.1 42.0
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 A 201
REMARK 800
REMARK 800 SITE_IDENTIFIER: AC2
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE CA A 202
REMARK 800
REMARK 800 SITE_IDENTIFIER: AC3
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE CA A 203
REMARK 800
REMARK 800 SITE_IDENTIFIER: AC4
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE CA A 204
REMARK 900
REMARK 900 RELATED ENTRIES
REMARK 900 RELATED ID: 3EWT RELATED DB: PDB
REMARK 900 CALMODULIN COMPLEX
REMARK 900 RELATED ID: 4DJC RELATED DB: PDB
REMARK 900 CALMODULIN COMPLEX
REMARK 900 RELATED ID: 25757 RELATED DB: BMRB
DBREF 2N6A A 5 146 UNP P62158 CALM_HUMAN 6 147
DBREF 2N6A A 155 171 UNP Q9UKL4 CXD2_HUMAN 276 292
SEQADV 2N6A HIS A -1 UNP P62158 EXPRESSION TAG
SEQADV 2N6A HIS A 0 UNP P62158 EXPRESSION TAG
SEQADV 2N6A HIS A 1 UNP P62158 EXPRESSION TAG
SEQADV 2N6A HIS A 2 UNP P62158 EXPRESSION TAG
SEQADV 2N6A HIS A 3 UNP P62158 EXPRESSION TAG
SEQADV 2N6A HIS A 4 UNP P62158 EXPRESSION TAG
SEQADV 2N6A GLY A 147 UNP P62158 LINKER
SEQADV 2N6A ALA A 148 UNP P62158 LINKER
SEQADV 2N6A SER A 149 UNP P62158 LINKER
SEQADV 2N6A THR A 150 UNP P62158 LINKER
SEQADV 2N6A ALA A 151 UNP P62158 LINKER
SEQADV 2N6A ALA A 152 UNP P62158 LINKER
SEQADV 2N6A GLY A 153 UNP P62158 LINKER
SEQADV 2N6A SER A 154 UNP P62158 LINKER
SEQRES 1 A 173 HIS HIS HIS HIS HIS HIS THR GLU GLU GLN ILE ALA GLU
SEQRES 2 A 173 PHE LYS GLU ALA PHE SER LEU PHE ASP LYS ASP GLY ASP
SEQRES 3 A 173 GLY THR ILE THR THR LYS GLU LEU GLY THR VAL MET ARG
SEQRES 4 A 173 SER LEU GLY GLN ASN PRO THR GLU ALA GLU LEU GLN ASP
SEQRES 5 A 173 MET ILE ASN GLU VAL ASP ALA ASP GLY ASN GLY THR ILE
SEQRES 6 A 173 ASP PHE PRO GLU PHE LEU THR MET MET ALA ARG LYS MET
SEQRES 7 A 173 LYS ASP THR ASP SER GLU GLU GLU ILE ARG GLU ALA PHE
SEQRES 8 A 173 ARG VAL PHE ASP LYS ASP GLY ASN GLY TYR ILE SER ALA
SEQRES 9 A 173 ALA GLU LEU ARG HIS VAL MET THR ASN LEU GLY GLU LYS
SEQRES 10 A 173 LEU THR ASP GLU GLU VAL ASP GLU MET ILE ARG GLU ALA
SEQRES 11 A 173 ASP ILE ASP GLY ASP GLY GLN VAL ASN TYR GLU GLU PHE
SEQRES 12 A 173 VAL GLN MET MET THR GLY ALA SER THR ALA ALA GLY SER
SEQRES 13 A 173 GLY TRP ARG LYS ILE LYS LEU ALA VAL ARG GLY ALA GLN
SEQRES 14 A 173 ALA LYS ARG LYS
HET CA A 201 1
HET CA A 202 1
HET CA A 203 1
HET CA A 204 1
HETNAM CA CALCIUM ION
FORMUL 2 CA 4(CA 2+)
HELIX 1 1 ILE A 9 LEU A 18 1 10
HELIX 2 2 THR A 28 SER A 38 1 11
HELIX 3 3 THR A 44 VAL A 55 1 12
HELIX 4 4 ASP A 64 ARG A 74 1 11
HELIX 5 5 LYS A 75 ASP A 78 5 4
HELIX 6 6 SER A 81 ASP A 93 1 13
HELIX 7 7 ALA A 102 GLY A 113 1 12
HELIX 8 8 THR A 117 ASP A 129 1 13
HELIX 9 9 TYR A 138 VAL A 142 1 5
HELIX 10 10 GLN A 143 THR A 146 5 4
HELIX 11 11 ALA A 152 ALA A 166 1 15
SHEET 1 A 2 TYR A 99 SER A 101 0
SHEET 2 A 2 GLN A 135 ASN A 137 -1 O VAL A 136 N ILE A 100
LINK OD1 ASP A 20 CA CA A 201 1555 1555 3.07
LINK OD2 ASP A 22 CA CA A 201 1555 1555 3.07
LINK OD2 ASP A 24 CA CA A 201 1555 1555 3.07
LINK O THR A 26 CA CA A 201 1555 1555 3.06
LINK OE1 GLU A 31 CA CA A 201 1555 1555 3.06
LINK OE2 GLU A 31 CA CA A 201 1555 1555 3.07
LINK OD1 ASP A 56 CA CA A 202 1555 1555 3.07
LINK OD1 ASP A 58 CA CA A 202 1555 1555 3.07
LINK OD1 ASN A 60 CA CA A 202 1555 1555 3.07
LINK O THR A 62 CA CA A 202 1555 1555 3.06
LINK OE2 GLU A 67 CA CA A 202 1555 1555 3.07
LINK OE1 GLU A 67 CA CA A 202 1555 1555 3.08
LINK OD1 ASP A 93 CA CA A 203 1555 1555 3.07
LINK OD2 ASP A 95 CA CA A 203 1555 1555 3.06
LINK OD1 ASN A 97 CA CA A 203 1555 1555 3.06
LINK O TYR A 99 CA CA A 203 1555 1555 3.07
LINK OE1 GLU A 104 CA CA A 203 1555 1555 3.06
LINK OE2 GLU A 104 CA CA A 203 1555 1555 3.07
LINK OD1 ASP A 129 CA CA A 204 1555 1555 3.06
LINK OD1 ASP A 131 CA CA A 204 1555 1555 3.07
LINK OD2 ASP A 133 CA CA A 204 1555 1555 3.06
LINK O GLN A 135 CA CA A 204 1555 1555 3.07
LINK OE2 GLU A 140 CA CA A 204 1555 1555 3.06
LINK OE1 GLU A 140 CA CA A 204 1555 1555 3.07
SITE 1 AC1 5 ASP A 20 ASP A 22 ASP A 24 THR A 26
SITE 2 AC1 5 GLU A 31
SITE 1 AC2 5 ASP A 56 ASP A 58 ASN A 60 THR A 62
SITE 2 AC2 5 GLU A 67
SITE 1 AC3 5 ASP A 93 ASP A 95 ASN A 97 TYR A 99
SITE 2 AC3 5 GLU A 104
SITE 1 AC4 5 ASP A 129 ASP A 131 ASP A 133 GLN A 135
SITE 2 AC4 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