HEADER METAL BINDING PROTEIN 17-FEB-10 2KUG
TITLE HALOTHANE BINDS TO DRUGGABLE SITES IN CALCIUM-CALMODULIN: SOLUTION
TITLE 2 STRUCTURE OF HALOTHANE-CAM N-TERMINAL DOMAIN
COMPND MOL_ID: 1;
COMPND 2 MOLECULE: CALMODULIN;
COMPND 3 CHAIN: A;
COMPND 4 FRAGMENT: N-TERMINAL DOMAIN EF-HANDS 1 AND 2;
COMPND 5 SYNONYM: CAM;
COMPND 6 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: CALM, CALM1, CALM2, CALM3, CALML2, CAM, CAM1, CAM2, CAM3,
SOURCE 6 CAMB, CAMC, CAMIII;
SOURCE 7 EXPRESSION_SYSTEM: ESCHERICHIA COLI;
SOURCE 8 EXPRESSION_SYSTEM_TAXID: 562;
SOURCE 9 EXPRESSION_SYSTEM_STRAIN: BL21(DE3)-PLYSS;
SOURCE 10 EXPRESSION_SYSTEM_VECTOR: PET 15B
KEYWDS CALMODULIN, CALCIUM BINDING, VOLATILE ANESTHETIC, HALOTHANE,
KEYWDS 2 CYTOSKELETON, ISOPEPTIDE BOND, METHYLATION, PHOSPHOPROTEIN, METAL
KEYWDS 3 BINDING PROTEIN
EXPDTA SOLUTION NMR
NUMMDL 5
AUTHOR N.JURANIC,S.MACURA,M.V.SIMEONOV,K.A.JONES,A.R.PENHEITER,T.J.HOCK,
AUTHOR 2 J.H.STREIFF
REVDAT 3 05-FEB-20 2KUG 1 REMARK
REVDAT 2 11-DEC-13 2KUG 1 JRNL VERSN
REVDAT 1 09-MAR-10 2KUG 0
JRNL AUTH N.O.JURANIC,K.A.JONES,A.R.PENHEITER,T.J.HOCK,J.H.STREIFF
JRNL TITL HALOTHANE BINDS TO DRUGGABLE SITES IN THE [CA2+]4-CALMODULIN
JRNL TITL 2 (CAM) COMPLEX, BUT DOES NOT INHIBIT [CA2+]4-CAM ACTIVATION
JRNL TITL 3 OF KINASE.
JRNL REF J. SERB. CHEM. SOC. V. 78 1655 2013
JRNL REFN ISSN 0352-5139
REMARK 2
REMARK 2 RESOLUTION. NOT APPLICABLE.
REMARK 3
REMARK 3 REFINEMENT.
REMARK 3 PROGRAM : X-PLOR MSI XPLOR 3.843, X-PLOR MSI XPLOR 3.843
REMARK 3 AUTHORS : BRUNGER (X-PLOR), BRUNGER (X-PLOR)
REMARK 3
REMARK 3 OTHER REFINEMENT REMARKS: 100 STEP STEEPEST DESCENT - FINAL
REMARK 3 REFINEMENT
REMARK 4
REMARK 4 2KUG COMPLIES WITH FORMAT V. 3.30, 13-JUL-11
REMARK 100
REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 03-MAR-10.
REMARK 100 THE DEPOSITION ID IS D_1000101586.
REMARK 210
REMARK 210 EXPERIMENTAL DETAILS
REMARK 210 EXPERIMENT TYPE : NMR
REMARK 210 TEMPERATURE (KELVIN) : 298
REMARK 210 PH : 7.2
REMARK 210 IONIC STRENGTH : 0.1
REMARK 210 PRESSURE : AMBIENT
REMARK 210 SAMPLE CONTENTS : 2 MM [U-99% 13C; U-99% 15N]
REMARK 210 CALMODULIN, 20 MM CALCIUM ION, 20 MM N-{[2-({[1-(4-
REMARK 210 CARBOXYBUTANOYL)AMINO]-2-PHENYLETHYL}-HYDROXYPHOSPHINYL)OXY]
REMARK 210 ACETYL}-2-PHENYLETHYLAMINE, 95% H2O/5% D2O; 2 MM [U-99% 15N]
REMARK 210 CALMODULIN, 20 MM CALCIUM ION, 20 MM N-{[2-({[1-(4-
REMARK 210 CARBOXYBUTANOYL)AMINO]-2-PHENYLETHYL}-HYDROXYPHOSPHINYL)OXY]
REMARK 210 ACETYL}-2-PHENYLETHYLAMINE, 95% H2O/5% D2O
REMARK 210
REMARK 210 NMR EXPERIMENTS CONDUCTED : 3D HNCO; 3D HNCA; 3D HNCACB; 3D
REMARK 210 HBHA(CO)NH; 3D H(CCO)NH; 3D HCCH-
REMARK 210 TOCSY; 3D 1H-15N NOESY; 3D 1H-
REMARK 210 13C NOESY; 3D HNHA; 3D HCCH-COSY
REMARK 210 SPECTROMETER FIELD STRENGTH : 700 MHZ; 500 MHZ
REMARK 210 SPECTROMETER MODEL : AVANCE
REMARK 210 SPECTROMETER MANUFACTURER : BRUKER
REMARK 210
REMARK 210 STRUCTURE DETERMINATION.
REMARK 210 SOFTWARE USED : NULL
REMARK 210 METHOD USED : SIMULATED ANNEALING, CHARMM22
REMARK 210 ENERGY MINIMIZATION
REMARK 210
REMARK 210 CONFORMERS, NUMBER CALCULATED : 50
REMARK 210 CONFORMERS, NUMBER SUBMITTED : 5
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 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 LYS A 75 84.74 -69.91
REMARK 500 2 LYS A 75 94.35 -64.43
REMARK 500 4 LYS A 75 90.71 -67.55
REMARK 500 5 LYS A 75 94.48 -64.38
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 990 CA
REMARK 620 N RES CSSEQI ATOM
REMARK 620 1 THR A 26 O
REMARK 620 2 ASP A 24 OD2 87.2
REMARK 620 3 GLU A 31 OE2 133.2 139.5
REMARK 620 4 GLU A 31 OE1 82.8 169.8 50.5
REMARK 620 5 ASP A 22 OD2 154.8 81.4 60.7 108.6
REMARK 620 6 ASP A 20 OD1 76.8 94.1 93.8 85.7 81.6
REMARK 620 7 ASP A 24 OD1 61.9 46.3 142.4 128.7 94.5 52.4
REMARK 620 8 ASP A 22 OD1 112.3 64.7 92.4 121.3 42.5 48.5 54.1
REMARK 620 N 1 2 3 4 5 6 7
REMARK 620
REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL
REMARK 620 CA A 991 CA
REMARK 620 N RES CSSEQI ATOM
REMARK 620 1 THR A 62 O
REMARK 620 2 ASP A 58 OD1 119.1
REMARK 620 3 GLU A 67 OE1 76.9 120.5
REMARK 620 4 ASP A 58 OD2 154.7 50.0 128.2
REMARK 620 5 GLU A 67 OE2 114.5 74.0 49.6 85.9
REMARK 620 6 ASP A 56 OD1 68.2 56.2 88.0 106.1 74.3
REMARK 620 7 ASN A 60 OD1 78.4 62.2 152.0 76.6 134.2 70.6
REMARK 620 N 1 2 3 4 5 6
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 990
REMARK 800
REMARK 800 SITE_IDENTIFIER: AC2
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE CA A 991
REMARK 800
REMARK 800 SITE_IDENTIFIER: AC3
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE HLT A 149
REMARK 900
REMARK 900 RELATED ENTRIES
REMARK 900 RELATED ID: 2KUH RELATED DB: PDB
REMARK 900 CAM O-TERMINAL DOMAIN STRUCTURE OF THE SAME COMPLEX
REMARK 900 RELATED ID: 16764 RELATED DB: BMRB
DBREF 2KUG A 1 76 UNP P62158 CALM_HUMAN 2 77
SEQRES 1 A 76 ALA ASP GLN LEU THR GLU GLU GLN ILE ALA GLU PHE LYS
SEQRES 2 A 76 GLU ALA PHE SER LEU PHE ASP LYS ASP GLY ASP GLY THR
SEQRES 3 A 76 ILE THR THR LYS GLU LEU GLY THR VAL MET ARG SER LEU
SEQRES 4 A 76 GLY GLN ASN PRO THR GLU ALA GLU LEU GLN ASP MET ILE
SEQRES 5 A 76 ASN GLU VAL ASP ALA ASP GLY ASN GLY THR ILE ASP PHE
SEQRES 6 A 76 PRO GLU PHE LEU THR MET MET ALA ARG LYS MET
HET CA A 990 1
HET CA A 991 1
HET HLT A 149 8
HETNAM CA CALCIUM ION
HETNAM HLT 2-BROMO-2-CHLORO-1,1,1-TRIFLUOROETHANE
FORMUL 2 CA 2(CA 2+)
FORMUL 4 HLT C2 H BR CL F3
HELIX 1 1 THR A 5 ASP A 20 1 16
HELIX 2 2 THR A 28 LEU A 39 1 12
HELIX 3 3 THR A 44 VAL A 55 1 12
HELIX 4 4 ASP A 64 LYS A 75 1 12
LINK O THR A 26 CA CA A 990 1555 1555 2.45
LINK O THR A 62 CA CA A 991 1555 1555 2.45
LINK OD2 ASP A 24 CA CA A 990 1555 1555 2.50
LINK OD1 ASP A 58 CA CA A 991 1555 1555 2.53
LINK OE2 GLU A 31 CA CA A 990 1555 1555 2.53
LINK OE1 GLU A 31 CA CA A 990 1555 1555 2.55
LINK OE1 GLU A 67 CA CA A 991 1555 1555 2.57
LINK OD2 ASP A 22 CA CA A 990 1555 1555 2.57
LINK OD2 ASP A 58 CA CA A 991 1555 1555 2.58
LINK OE2 GLU A 67 CA CA A 991 1555 1555 2.59
LINK OD1 ASP A 56 CA CA A 991 1555 1555 2.61
LINK OD1 ASN A 60 CA CA A 991 1555 1555 2.62
LINK OD1 ASP A 20 CA CA A 990 1555 1555 2.72
LINK OD1 ASP A 24 CA CA A 990 1555 1555 2.90
LINK OD1 ASP A 22 CA CA A 990 1555 1555 3.15
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 3 MET A 36 MET A 51 MET A 71
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