HEADER FLUORESCENT PROTEIN 18-SEP-08 3EK7
TITLE CALCIUM-SATURATED GCAMP2 DIMER
COMPND MOL_ID: 1;
COMPND 2 MOLECULE: MYOSIN LIGHT CHAIN KINASE, GREEN FLUORESCENT PROTEIN,
COMPND 3 CALMODULIN CHIMERA;
COMPND 4 CHAIN: A;
COMPND 5 ENGINEERED: YES
SOURCE MOL_ID: 1;
SOURCE 2 ORGANISM_SCIENTIFIC: ARTIFICIAL GENE, AEQUOREA VICTORIA, RATTUS
SOURCE 3 NORVEGICUS;
SOURCE 4 ORGANISM_COMMON: JELLYFISH, RAT;
SOURCE 5 ORGANISM_TAXID: 32630, 6100, 10116;
SOURCE 6 GENE: GFP, CALM1, CALM, CAM, CAM1, CAMI;
SOURCE 7 EXPRESSION_SYSTEM: ESCHERICHIA COLI;
SOURCE 8 EXPRESSION_SYSTEM_TAXID: 469008;
SOURCE 9 EXPRESSION_SYSTEM_STRAIN: BL21(DE3);
SOURCE 10 EXPRESSION_SYSTEM_VECTOR_TYPE: PLASMID;
SOURCE 11 EXPRESSION_SYSTEM_PLASMID: PRSETA
KEYWDS GECI, GCAMP2, CPEGFP, CALMODULIN, M13 PEPTIDE, METHYLATION,
KEYWDS 2 PHOSPHOPROTEIN, SIGNALING PROTEIN, FLUORESCENT PROTEIN
EXPDTA X-RAY DIFFRACTION
AUTHOR J.AKERBOOM,J.D.VELEZ RIVERA,L.L.LOOGER,E.R.SCHREITER
REVDAT 8 15-NOV-23 3EK7 1 REMARK
REVDAT 7 30-AUG-23 3EK7 1 REMARK LINK
REVDAT 6 25-OCT-17 3EK7 1 REMARK
REVDAT 5 21-JUN-17 3EK7 1 SOURCE DBREF SEQADV MODRES
REVDAT 4 31-OCT-12 3EK7 1 DBREF VERSN
REVDAT 3 10-MAR-09 3EK7 1 JRNL
REVDAT 2 30-DEC-08 3EK7 1 JRNL
REVDAT 1 16-DEC-08 3EK7 0
JRNL AUTH J.AKERBOOM,J.D.RIVERA,M.M.GUILBE,E.C.MALAVE,H.H.HERNANDEZ,
JRNL AUTH 2 L.TIAN,S.A.HIRES,J.S.MARVIN,L.L.LOOGER,E.R.SCHREITER
JRNL TITL CRYSTAL STRUCTURES OF THE GCAMP CALCIUM SENSOR REVEAL THE
JRNL TITL 2 MECHANISM OF FLUORESCENCE SIGNAL CHANGE AND AID RATIONAL
JRNL TITL 3 DESIGN
JRNL REF J.BIOL.CHEM. V. 284 6455 2009
JRNL REFN ISSN 0021-9258
JRNL PMID 19098007
JRNL DOI 10.1074/JBC.M807657200
REMARK 1
REMARK 1 REFERENCE 1
REMARK 1 AUTH M.M RODRIGUEZ GUILBE,E.C.ALFARO MALAVE,J.AKERBOOM,
REMARK 1 AUTH 2 J.S.MARVIN,L.L.LOOGER,E.R.SCHREITER
REMARK 1 TITL CRYSTALLIZATION AND PRELIMINARY X-RAY CHARACTERIZATION OF
REMARK 1 TITL 2 THE GENETICALLY ENCODED FLUORESCENT CALCIUM INDICATOR
REMARK 1 TITL 3 PROTEIN GCAMP2
REMARK 1 REF ACTA CRYSTALLOGR.,SECT.F V. 64 629 2008
REMARK 1 REFN ESSN 1744-3091
REMARK 2
REMARK 2 RESOLUTION. 1.85 ANGSTROMS.
REMARK 3
REMARK 3 REFINEMENT.
REMARK 3 PROGRAM : REFMAC 5.2.0019
REMARK 3 AUTHORS : MURSHUDOV,SKUBAK,LEBEDEV,PANNU,STEINER,
REMARK 3 : NICHOLLS,WINN,LONG,VAGIN
REMARK 3
REMARK 3 REFINEMENT TARGET : MAXIMUM LIKELIHOOD
REMARK 3
REMARK 3 DATA USED IN REFINEMENT.
REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 1.85
REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 20.67
REMARK 3 DATA CUTOFF (SIGMA(F)) : NULL
REMARK 3 COMPLETENESS FOR RANGE (%) : 98.5
REMARK 3 NUMBER OF REFLECTIONS : 32223
REMARK 3
REMARK 3 FIT TO DATA USED IN REFINEMENT.
REMARK 3 CROSS-VALIDATION METHOD : THROUGHOUT
REMARK 3 FREE R VALUE TEST SET SELECTION : RANDOM
REMARK 3 R VALUE (WORKING + TEST SET) : 0.191
REMARK 3 R VALUE (WORKING SET) : 0.189
REMARK 3 FREE R VALUE : 0.241
REMARK 3 FREE R VALUE TEST SET SIZE (%) : 5.100
REMARK 3 FREE R VALUE TEST SET COUNT : 1714
REMARK 3
REMARK 3 FIT IN THE HIGHEST RESOLUTION BIN.
REMARK 3 TOTAL NUMBER OF BINS USED : 20
REMARK 3 BIN RESOLUTION RANGE HIGH (A) : 1.85
REMARK 3 BIN RESOLUTION RANGE LOW (A) : 1.90
REMARK 3 REFLECTION IN BIN (WORKING SET) : 2378
REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 97.24
REMARK 3 BIN R VALUE (WORKING SET) : 0.2070
REMARK 3 BIN FREE R VALUE SET COUNT : 121
REMARK 3 BIN FREE R VALUE : 0.2850
REMARK 3
REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.
REMARK 3 PROTEIN ATOMS : 3105
REMARK 3 NUCLEIC ACID ATOMS : 0
REMARK 3 HETEROGEN ATOMS : 4
REMARK 3 SOLVENT ATOMS : 198
REMARK 3
REMARK 3 B VALUES.
REMARK 3 FROM WILSON PLOT (A**2) : NULL
REMARK 3 MEAN B VALUE (OVERALL, A**2) : 20.84
REMARK 3 OVERALL ANISOTROPIC B VALUE.
REMARK 3 B11 (A**2) : -0.09000
REMARK 3 B22 (A**2) : 0.60000
REMARK 3 B33 (A**2) : -0.73000
REMARK 3 B12 (A**2) : 0.00000
REMARK 3 B13 (A**2) : -0.57000
REMARK 3 B23 (A**2) : 0.00000
REMARK 3
REMARK 3 ESTIMATED OVERALL COORDINATE ERROR.
REMARK 3 ESU BASED ON R VALUE (A): 0.150
REMARK 3 ESU BASED ON FREE R VALUE (A): 0.146
REMARK 3 ESU BASED ON MAXIMUM LIKELIHOOD (A): 0.095
REMARK 3 ESU FOR B VALUES BASED ON MAXIMUM LIKELIHOOD (A**2): 3.064
REMARK 3
REMARK 3 CORRELATION COEFFICIENTS.
REMARK 3 CORRELATION COEFFICIENT FO-FC : 0.951
REMARK 3 CORRELATION COEFFICIENT FO-FC FREE : 0.923
REMARK 3
REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES COUNT RMS WEIGHT
REMARK 3 BOND LENGTHS REFINED ATOMS (A): 3191 ; 0.018 ; 0.022
REMARK 3 BOND LENGTHS OTHERS (A): 2172 ; 0.002 ; 0.020
REMARK 3 BOND ANGLES REFINED ATOMS (DEGREES): 4305 ; 1.735 ; 1.968
REMARK 3 BOND ANGLES OTHERS (DEGREES): 5311 ; 0.986 ; 3.000
REMARK 3 TORSION ANGLES, PERIOD 1 (DEGREES): 393 ; 6.199 ; 5.000
REMARK 3 TORSION ANGLES, PERIOD 2 (DEGREES): 160 ;34.391 ;25.312
REMARK 3 TORSION ANGLES, PERIOD 3 (DEGREES): 579 ;15.393 ;15.000
REMARK 3 TORSION ANGLES, PERIOD 4 (DEGREES): 16 ;17.070 ;15.000
REMARK 3 CHIRAL-CENTER RESTRAINTS (A**3): 467 ; 0.097 ; 0.200
REMARK 3 GENERAL PLANES REFINED ATOMS (A): 3582 ; 0.007 ; 0.020
REMARK 3 GENERAL PLANES OTHERS (A): 628 ; 0.002 ; 0.020
REMARK 3 NON-BONDED CONTACTS REFINED ATOMS (A): 648 ; 0.218 ; 0.200
REMARK 3 NON-BONDED CONTACTS OTHERS (A): 2115 ; 0.199 ; 0.200
REMARK 3 NON-BONDED TORSION REFINED ATOMS (A): 1506 ; 0.178 ; 0.200
REMARK 3 NON-BONDED TORSION OTHERS (A): 1719 ; 0.111 ; 0.200
REMARK 3 H-BOND (X...Y) REFINED ATOMS (A): 161 ; 0.150 ; 0.200
REMARK 3 H-BOND (X...Y) OTHERS (A): NULL ; NULL ; NULL
REMARK 3 POTENTIAL METAL-ION REFINED ATOMS (A): 12 ; 0.154 ; 0.200
REMARK 3 POTENTIAL METAL-ION OTHERS (A): NULL ; NULL ; NULL
REMARK 3 SYMMETRY VDW REFINED ATOMS (A): 18 ; 0.148 ; 0.200
REMARK 3 SYMMETRY VDW OTHERS (A): 63 ; 0.274 ; 0.200
REMARK 3 SYMMETRY H-BOND REFINED ATOMS (A): 18 ; 0.527 ; 0.200
REMARK 3 SYMMETRY H-BOND OTHERS (A): NULL ; NULL ; NULL
REMARK 3 SYMMETRY METAL-ION REFINED ATOMS (A): NULL ; NULL ; NULL
REMARK 3 SYMMETRY METAL-ION OTHERS (A): NULL ; NULL ; NULL
REMARK 3
REMARK 3 ISOTROPIC THERMAL FACTOR RESTRAINTS. COUNT RMS WEIGHT
REMARK 3 MAIN-CHAIN BOND REFINED ATOMS (A**2): 2027 ; 1.404 ; 1.500
REMARK 3 MAIN-CHAIN BOND OTHER ATOMS (A**2): 808 ; 0.316 ; 1.500
REMARK 3 MAIN-CHAIN ANGLE REFINED ATOMS (A**2): 3134 ; 1.991 ; 2.000
REMARK 3 MAIN-CHAIN ANGLE OTHER ATOMS (A**2): NULL ; NULL ; NULL
REMARK 3 SIDE-CHAIN BOND REFINED ATOMS (A**2): 1344 ; 3.200 ; 3.000
REMARK 3 SIDE-CHAIN BOND OTHER ATOMS (A**2): NULL ; NULL ; NULL
REMARK 3 SIDE-CHAIN ANGLE REFINED ATOMS (A**2): 1170 ; 4.699 ; 4.500
REMARK 3 SIDE-CHAIN ANGLE OTHER ATOMS (A**2): NULL ; NULL ; NULL
REMARK 3 LONG RANGE B REFINED ATOMS (A**2): NULL ; NULL ; NULL
REMARK 3 LONG RANGE B OTHER ATOMS (A**2): NULL ; NULL ; NULL
REMARK 3
REMARK 3 ANISOTROPIC THERMAL FACTOR RESTRAINTS. COUNT RMS WEIGHT
REMARK 3 RIGID-BOND RESTRAINTS (A**2): NULL ; NULL ; NULL
REMARK 3 SPHERICITY; FREE ATOMS (A**2): NULL ; NULL ; NULL
REMARK 3 SPHERICITY; BONDED ATOMS (A**2): NULL ; NULL ; NULL
REMARK 3
REMARK 3 NCS RESTRAINTS STATISTICS
REMARK 3 NUMBER OF DIFFERENT NCS GROUPS : NULL
REMARK 3
REMARK 3 TLS DETAILS
REMARK 3 NUMBER OF TLS GROUPS : NULL
REMARK 3
REMARK 3 BULK SOLVENT MODELLING.
REMARK 3 METHOD USED : MASK
REMARK 3 PARAMETERS FOR MASK CALCULATION
REMARK 3 VDW PROBE RADIUS : 1.20
REMARK 3 ION PROBE RADIUS : 0.80
REMARK 3 SHRINKAGE RADIUS : 0.80
REMARK 3
REMARK 3 OTHER REFINEMENT REMARKS: HYDROGENS HAVE BEEN ADDED IN THE RIDING
REMARK 3 POSITIONS
REMARK 4
REMARK 4 3EK7 COMPLIES WITH FORMAT V. 3.30, 13-JUL-11
REMARK 100
REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 22-SEP-08.
REMARK 100 THE DEPOSITION ID IS D_1000049425.
REMARK 200
REMARK 200 EXPERIMENTAL DETAILS
REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION
REMARK 200 DATE OF DATA COLLECTION : 28-JUL-08
REMARK 200 TEMPERATURE (KELVIN) : 100
REMARK 200 PH : 8.5
REMARK 200 NUMBER OF CRYSTALS USED : 1
REMARK 200
REMARK 200 SYNCHROTRON (Y/N) : Y
REMARK 200 RADIATION SOURCE : APS
REMARK 200 BEAMLINE : 31-ID
REMARK 200 X-RAY GENERATOR MODEL : NULL
REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M
REMARK 200 WAVELENGTH OR RANGE (A) : 0.9793
REMARK 200 MONOCHROMATOR : DIAMOND(1,1,1)
REMARK 200 OPTICS : NULL
REMARK 200
REMARK 200 DETECTOR TYPE : CCD
REMARK 200 DETECTOR MANUFACTURER : MAR CCD 165 MM
REMARK 200 INTENSITY-INTEGRATION SOFTWARE : DENZO
REMARK 200 DATA SCALING SOFTWARE : SCALA, SCALEPACK
REMARK 200
REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 36788
REMARK 200 RESOLUTION RANGE HIGH (A) : 1.850
REMARK 200 RESOLUTION RANGE LOW (A) : 67.730
REMARK 200 REJECTION CRITERIA (SIGMA(I)) : 0.000
REMARK 200
REMARK 200 OVERALL.
REMARK 200 COMPLETENESS FOR RANGE (%) : 98.5
REMARK 200 DATA REDUNDANCY : 7.500
REMARK 200 R MERGE (I) : NULL
REMARK 200 R SYM (I) : 0.07600
REMARK 200 <I/SIGMA(I)> FOR THE DATA SET : NULL
REMARK 200
REMARK 200 IN THE HIGHEST RESOLUTION SHELL.
REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 1.85
REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 1.90
REMARK 200 COMPLETENESS FOR SHELL (%) : 97.6
REMARK 200 DATA REDUNDANCY IN SHELL : 7.60
REMARK 200 R MERGE FOR SHELL (I) : NULL
REMARK 200 R SYM FOR SHELL (I) : 0.42000
REMARK 200 <I/SIGMA(I)> FOR SHELL : 5.100
REMARK 200
REMARK 200 DIFFRACTION PROTOCOL: SINGLE WAVELENGTH
REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: MOLECULAR REPLACEMENT
REMARK 200 SOFTWARE USED: PHASER
REMARK 200 STARTING MODEL: PDB ENTRIES 1EMA AND 1CDL
REMARK 200
REMARK 200 REMARK: NULL
REMARK 280
REMARK 280 CRYSTAL
REMARK 280 SOLVENT CONTENT, VS (%): 38.06
REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 1.99
REMARK 280
REMARK 280 CRYSTALLIZATION CONDITIONS: 0.2 M LITHIUM SULFATE MONOHYDRATE, 0.1
REMARK 280 M TRIS-HCL PH 8.5, 30%(W/V) POLYETHYLENE GLYCOL 4000, VAPOR
REMARK 280 DIFFUSION, HANGING DROP, TEMPERATURE 298K
REMARK 290
REMARK 290 CRYSTALLOGRAPHIC SYMMETRY
REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: C 1 2 1
REMARK 290
REMARK 290 SYMOP SYMMETRY
REMARK 290 NNNMMM OPERATOR
REMARK 290 1555 X,Y,Z
REMARK 290 2555 -X,Y,-Z
REMARK 290 3555 X+1/2,Y+1/2,Z
REMARK 290 4555 -X+1/2,Y+1/2,-Z
REMARK 290
REMARK 290 WHERE NNN -> OPERATOR NUMBER
REMARK 290 MMM -> TRANSLATION VECTOR
REMARK 290
REMARK 290 CRYSTALLOGRAPHIC SYMMETRY TRANSFORMATIONS
REMARK 290 THE FOLLOWING TRANSFORMATIONS OPERATE ON THE ATOM/HETATM
REMARK 290 RECORDS IN THIS ENTRY TO PRODUCE CRYSTALLOGRAPHICALLY
REMARK 290 RELATED MOLECULES.
REMARK 290 SMTRY1 1 1.000000 0.000000 0.000000 0.00000
REMARK 290 SMTRY2 1 0.000000 1.000000 0.000000 0.00000
REMARK 290 SMTRY3 1 0.000000 0.000000 1.000000 0.00000
REMARK 290 SMTRY1 2 -1.000000 0.000000 0.000000 0.00000
REMARK 290 SMTRY2 2 0.000000 1.000000 0.000000 0.00000
REMARK 290 SMTRY3 2 0.000000 0.000000 -1.000000 0.00000
REMARK 290 SMTRY1 3 1.000000 0.000000 0.000000 63.06550
REMARK 290 SMTRY2 3 0.000000 1.000000 0.000000 23.65500
REMARK 290 SMTRY3 3 0.000000 0.000000 1.000000 0.00000
REMARK 290 SMTRY1 4 -1.000000 0.000000 0.000000 63.06550
REMARK 290 SMTRY2 4 0.000000 1.000000 0.000000 23.65500
REMARK 290 SMTRY3 4 0.000000 0.000000 -1.000000 0.00000
REMARK 290
REMARK 290 REMARK: NULL
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 SOFTWARE DETERMINED QUATERNARY STRUCTURE: DIMERIC
REMARK 350 SOFTWARE USED: PISA
REMARK 350 TOTAL BURIED SURFACE AREA: 7120 ANGSTROM**2
REMARK 350 SURFACE AREA OF THE COMPLEX: 35270 ANGSTROM**2
REMARK 350 CHANGE IN SOLVENT FREE ENERGY: -153.0 KCAL/MOL
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 350 BIOMT1 2 -1.000000 0.000000 0.000000 0.00000
REMARK 350 BIOMT2 2 0.000000 1.000000 0.000000 0.00000
REMARK 350 BIOMT3 2 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. (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN
REMARK 465 IDENTIFIER; SSSEQ=SEQUENCE NUMBER; I=INSERTION CODE.)
REMARK 465
REMARK 465 M RES C SSSEQI
REMARK 465 MET A 1
REMARK 465 ARG A 2
REMARK 465 GLY A 3
REMARK 465 SER A 4
REMARK 465 HIS A 5
REMARK 465 HIS A 6
REMARK 465 HIS A 7
REMARK 465 HIS A 8
REMARK 465 HIS A 9
REMARK 465 HIS A 10
REMARK 465 GLY A 11
REMARK 465 MET A 12
REMARK 465 ALA A 13
REMARK 465 SER A 14
REMARK 465 MET A 15
REMARK 465 THR A 16
REMARK 465 GLY A 17
REMARK 465 GLY A 18
REMARK 465 GLN A 19
REMARK 465 GLN A 20
REMARK 465 MET A 21
REMARK 465 GLY A 22
REMARK 465 ARG A 23
REMARK 465 ASP A 24
REMARK 465 LEU A 25
REMARK 465 TYR A 26
REMARK 465 ASP A 27
REMARK 465 ASP A 28
REMARK 465 ASP A 29
REMARK 465 ASP A 30
REMARK 465 LYS A 31
REMARK 465 ASP A 32
REMARK 465 LEU A 33
REMARK 465 ALA A 34
REMARK 465 THR A 35
REMARK 465 MET A 36
REMARK 465 VAL A 37
REMARK 465 ASP A 38
REMARK 465 GLY A 145
REMARK 465 MET A 146
REMARK 465 ASP A 147
REMARK 465 GLU A 148
REMARK 465 LEU A 149
REMARK 465 TYR A 150
REMARK 465 LYS A 151
REMARK 465 GLY A 152
REMARK 465 GLY A 153
REMARK 465 THR A 154
REMARK 465 GLY A 155
REMARK 465 GLY A 156
REMARK 465 SER A 157
REMARK 465 MET A 158
REMARK 465 VAL A 159
REMARK 465 THR A 303
REMARK 465 ARG A 304
REMARK 465 ASP A 305
REMARK 465 GLN A 306
REMARK 465 ALA A 450
REMARK 465 LYS A 451
REMARK 500
REMARK 500 GEOMETRY AND STEREOCHEMISTRY
REMARK 500 SUBTOPIC: CLOSE CONTACTS IN SAME ASYMMETRIC UNIT
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 HOH A 652 O HOH A 653 1.45
REMARK 500 O HOH A 631 O HOH A 648 1.68
REMARK 500 O HOH A 642 O HOH A 651 1.77
REMARK 500 OD2 ASP A 194 O HOH A 611 1.95
REMARK 500
REMARK 500 REMARK: NULL
REMARK 500
REMARK 500 GEOMETRY AND STEREOCHEMISTRY
REMARK 500 SUBTOPIC: CLOSE CONTACTS
REMARK 500
REMARK 500 THE FOLLOWING ATOMS THAT ARE RELATED BY CRYSTALLOGRAPHIC
REMARK 500 SYMMETRY ARE IN CLOSE CONTACT. AN ATOM LOCATED WITHIN 0.15
REMARK 500 ANGSTROMS OF A SYMMETRY RELATED ATOM IS ASSUMED TO BE ON A
REMARK 500 SPECIAL POSITION AND IS, THEREFORE, LISTED IN REMARK 375
REMARK 500 INSTEAD OF REMARK 500. ATOMS WITH NON-BLANK ALTERNATE
REMARK 500 LOCATION INDICATORS ARE NOT INCLUDED IN THE CALCULATIONS.
REMARK 500
REMARK 500 DISTANCE CUTOFF:
REMARK 500 2.2 ANGSTROMS FOR CONTACTS NOT INVOLVING HYDROGEN ATOMS
REMARK 500 1.6 ANGSTROMS FOR CONTACTS INVOLVING HYDROGEN ATOMS
REMARK 500
REMARK 500 ATM1 RES C SSEQI ATM2 RES C SSEQI SSYMOP DISTANCE
REMARK 500 O HOH A 627 O HOH A 643 1545 1.76
REMARK 500
REMARK 500 REMARK: NULL
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 PROTEIN: ENGH AND HUBER, 1999
REMARK 500 EXPECTED VALUES NUCLEIC ACID: CLOWNEY ET AL 1996
REMARK 500
REMARK 500 M RES CSSEQI ATM1 ATM2 ATM3
REMARK 500 ARG A 81 NE - CZ - NH2 ANGL. DEV. = -3.4 DEGREES
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 ARG A 377 78.38 -63.46
REMARK 500 LYS A 378 144.03 -38.63
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 452 CA
REMARK 620 N RES CSSEQI ATOM
REMARK 620 1 ASP A 323 OD1
REMARK 620 2 ASP A 325 OD1 81.9
REMARK 620 3 ASP A 327 OD1 84.1 75.8
REMARK 620 4 THR A 329 O 87.8 156.5 82.2
REMARK 620 5 GLU A 334 OE1 98.5 74.6 149.6 128.1
REMARK 620 6 GLU A 334 OE2 111.0 126.3 153.5 77.1 52.5
REMARK 620 7 HOH A 507 O 159.7 78.4 86.5 108.7 80.8 84.8
REMARK 620 N 1 2 3 4 5 6
REMARK 620
REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL
REMARK 620 CA A 453 CA
REMARK 620 N RES CSSEQI ATOM
REMARK 620 1 ASP A 359 OD2
REMARK 620 2 ASP A 361 OD1 80.3
REMARK 620 3 ASN A 363 OD1 87.8 74.4
REMARK 620 4 THR A 365 O 87.2 153.3 81.6
REMARK 620 5 GLU A 370 OE1 108.2 126.2 155.0 80.1
REMARK 620 6 GLU A 370 OE2 88.1 76.9 151.3 126.5 51.3
REMARK 620 7 HOH A 518 O 159.4 79.1 86.6 111.5 84.3 87.4
REMARK 620 N 1 2 3 4 5 6
REMARK 620
REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL
REMARK 620 CA A 454 CA
REMARK 620 N RES CSSEQI ATOM
REMARK 620 1 ASP A 396 OD2
REMARK 620 2 ASP A 398 OD2 80.5
REMARK 620 3 ASN A 400 OD1 86.5 75.0
REMARK 620 4 TYR A 402 O 89.0 157.5 84.6
REMARK 620 5 GLU A 407 OE1 93.5 70.2 144.6 130.8
REMARK 620 6 GLU A 407 OE2 100.2 121.0 163.4 80.4 50.8
REMARK 620 7 HOH A 550 O 167.3 103.5 82.9 82.9 99.2 88.2
REMARK 620 N 1 2 3 4 5 6
REMARK 620
REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL
REMARK 620 CA A 455 CA
REMARK 620 N RES CSSEQI ATOM
REMARK 620 1 ASP A 432 OD1
REMARK 620 2 ASP A 434 OD2 84.3
REMARK 620 3 ASP A 436 OD1 81.9 80.7
REMARK 620 4 GLN A 438 O 83.0 158.1 79.9
REMARK 620 5 GLU A 443 OE1 112.3 122.3 152.8 79.1
REMARK 620 6 GLU A 443 OE2 94.7 73.8 154.5 124.9 50.9
REMARK 620 7 HOH A 525 O 162.7 90.2 81.0 96.8 84.6 99.5
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 452
REMARK 800
REMARK 800 SITE_IDENTIFIER: AC2
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE CA A 453
REMARK 800
REMARK 800 SITE_IDENTIFIER: AC3
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE CA A 454
REMARK 800
REMARK 800 SITE_IDENTIFIER: AC4
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE CA A 455
REMARK 900
REMARK 900 RELATED ENTRIES
REMARK 900 RELATED ID: 3EK4 RELATED DB: PDB
REMARK 900 CALCIUM-SATURATED GCAMP2 MONOMER
REMARK 900 RELATED ID: 3EK8 RELATED DB: PDB
REMARK 900 CALCIUM-SATURATED GCAMP2 T116V/G174R MONOMER
REMARK 900 RELATED ID: 3EKH RELATED DB: PDB
REMARK 900 CALCIUM-SATURATED GCAMP2 T116V/K387W MONOMER
REMARK 900 RELATED ID: 3EKJ RELATED DB: PDB
REMARK 999
REMARK 999 SEQUENCE
REMARK 999 RESIDUE SER65 HAS BEEN MUTATED TO GLY. RESIDUES GLY65, TYR66, AND
REMARK 999 GLY67 CONSTITUTE THE CHROMOPHORE CRO. THE AUTHOR STATES THAT ANY
REMARK 999 DIFFERENCES WITH DATABASE SEQUENCE LIKELY REPRESENT MUTATIONS
REMARK 999 INTRODUCED DURING VARIOUS ENGINEERING STEPS.
DBREF 3EK7 A 1 61 PDB 3EK7 3EK7 1 61
DBREF 3EK7 A 62 151 UNP P42212 GFP_AEQVI 149 238
DBREF 3EK7 A 160 302 UNP P42212 GFP_AEQVI 2 144
DBREF 3EK7 A 305 451 UNP P0DP29 CALM1_RAT 3 149
SEQADV 3EK7 ALA A 76 UNP P42212 VAL 163 SEE REMARK 999
SEQADV 3EK7 GLY A 88 UNP P42212 SER 175 SEE REMARK 999
SEQADV 3EK7 TYR A 93 UNP P42212 ASP 180 SEE REMARK 999
SEQADV 3EK7 LYS A 119 UNP P42212 ALA 206 SEE REMARK 999
SEQADV 3EK7 LEU A 144 UNP P42212 HIS 231 SEE REMARK 999
SEQADV 3EK7 GLY A 152 UNP P42212 LINKER
SEQADV 3EK7 GLY A 153 UNP P42212 LINKER
SEQADV 3EK7 THR A 154 UNP P42212 LINKER
SEQADV 3EK7 GLY A 155 UNP P42212 LINKER
SEQADV 3EK7 GLY A 156 UNP P42212 LINKER
SEQADV 3EK7 SER A 157 UNP P42212 LINKER
SEQADV 3EK7 MET A 158 UNP P42212 LINKER
SEQADV 3EK7 VAL A 159 UNP P42212 LINKER
SEQADV 3EK7 LEU A 222 UNP P42212 PHE 64 SEE REMARK 999
SEQADV 3EK7 CRO A 224 UNP P42212 SER 65 CHROMOPHORE
SEQADV 3EK7 A UNP P42212 TYR 66 CHROMOPHORE
SEQADV 3EK7 A UNP P42212 GLY 67 CHROMOPHORE
SEQADV 3EK7 ILE A 251 UNP P42212 VAL 93 SEE REMARK 999
SEQADV 3EK7 THR A 303 UNP P42212 LINKER
SEQADV 3EK7 ARG A 304 UNP P42212 LINKER
SEQRES 1 A 449 MET ARG GLY SER HIS HIS HIS HIS HIS HIS GLY MET ALA
SEQRES 2 A 449 SER MET THR GLY GLY GLN GLN MET GLY ARG ASP LEU TYR
SEQRES 3 A 449 ASP ASP ASP ASP LYS ASP LEU ALA THR MET VAL ASP SER
SEQRES 4 A 449 SER ARG ARG LYS TRP ASN LYS THR GLY HIS ALA VAL ARG
SEQRES 5 A 449 ALA ILE GLY ARG LEU SER SER LEU GLU ASN VAL TYR ILE
SEQRES 6 A 449 MET ALA ASP LYS GLN LYS ASN GLY ILE LYS ALA ASN PHE
SEQRES 7 A 449 LYS ILE ARG HIS ASN ILE GLU ASP GLY GLY VAL GLN LEU
SEQRES 8 A 449 ALA TYR HIS TYR GLN GLN ASN THR PRO ILE GLY ASP GLY
SEQRES 9 A 449 PRO VAL LEU LEU PRO ASP ASN HIS TYR LEU SER THR GLN
SEQRES 10 A 449 SER LYS LEU SER LYS ASP PRO ASN GLU LYS ARG ASP HIS
SEQRES 11 A 449 MET VAL LEU LEU GLU PHE VAL THR ALA ALA GLY ILE THR
SEQRES 12 A 449 LEU GLY MET ASP GLU LEU TYR LYS GLY GLY THR GLY GLY
SEQRES 13 A 449 SER MET VAL SER LYS GLY GLU GLU LEU PHE THR GLY VAL
SEQRES 14 A 449 VAL PRO ILE LEU VAL GLU LEU ASP GLY ASP VAL ASN GLY
SEQRES 15 A 449 HIS LYS PHE SER VAL SER GLY GLU GLY GLU GLY ASP ALA
SEQRES 16 A 449 THR TYR GLY LYS LEU THR LEU LYS PHE ILE CYS THR THR
SEQRES 17 A 449 GLY LYS LEU PRO VAL PRO TRP PRO THR LEU VAL THR THR
SEQRES 18 A 449 LEU CRO VAL GLN CYS PHE SER ARG TYR PRO ASP HIS MET
SEQRES 19 A 449 LYS GLN HIS ASP PHE PHE LYS SER ALA MET PRO GLU GLY
SEQRES 20 A 449 TYR ILE GLN GLU ARG THR ILE PHE PHE LYS ASP ASP GLY
SEQRES 21 A 449 ASN TYR LYS THR ARG ALA GLU VAL LYS PHE GLU GLY ASP
SEQRES 22 A 449 THR LEU VAL ASN ARG ILE GLU LEU LYS GLY ILE ASP PHE
SEQRES 23 A 449 LYS GLU ASP GLY ASN ILE LEU GLY HIS LYS LEU GLU TYR
SEQRES 24 A 449 ASN THR ARG ASP GLN LEU THR GLU GLU GLN ILE ALA GLU
SEQRES 25 A 449 PHE LYS GLU ALA PHE SER LEU PHE ASP LYS ASP GLY ASP
SEQRES 26 A 449 GLY THR ILE THR THR LYS GLU LEU GLY THR VAL MET ARG
SEQRES 27 A 449 SER LEU GLY GLN ASN PRO THR GLU ALA GLU LEU GLN ASP
SEQRES 28 A 449 MET ILE ASN GLU VAL ASP ALA ASP GLY ASN GLY THR ILE
SEQRES 29 A 449 ASP PHE PRO GLU PHE LEU THR MET MET ALA ARG LYS MET
SEQRES 30 A 449 LYS ASP THR ASP SER GLU GLU GLU ILE ARG GLU ALA PHE
SEQRES 31 A 449 ARG VAL PHE ASP LYS ASP GLY ASN GLY TYR ILE SER ALA
SEQRES 32 A 449 ALA GLU LEU ARG HIS VAL MET THR ASN LEU GLY GLU LYS
SEQRES 33 A 449 LEU THR ASP GLU GLU VAL ASP GLU MET ILE ARG GLU ALA
SEQRES 34 A 449 ASP ILE ASP GLY ASP GLY GLN VAL ASN TYR GLU GLU PHE
SEQRES 35 A 449 VAL GLN MET MET THR ALA LYS
MODRES 3EK7 CRO A 224 SER CHROMOPHORE
HET CRO A 224 22
HET CA A 452 1
HET CA A 453 1
HET CA A 454 1
HET CA A 455 1
HETNAM CRO {2-[(1R,2R)-1-AMINO-2-HYDROXYPROPYL]-4-(4-
HETNAM 2 CRO HYDROXYBENZYLIDENE)-5-OXO-4,5-DIHYDRO-1H-IMIDAZOL-1-
HETNAM 3 CRO YL}ACETIC ACID
HETNAM CA CALCIUM ION
HETSYN CRO PEPTIDE DERIVED CHROMOPHORE
FORMUL 1 CRO C15 H17 N3 O5
FORMUL 2 CA 4(CA 2+)
FORMUL 6 HOH *198(H2 O)
HELIX 1 1 SER A 39 SER A 59 1 21
HELIX 2 2 LYS A 69 ASN A 72 5 4
HELIX 3 3 SER A 160 LEU A 165 1 6
HELIX 4 4 PRO A 214 VAL A 219 5 6
HELIX 5 5 VAL A 226 SER A 230 5 5
HELIX 6 6 PRO A 233 HIS A 239 5 7
HELIX 7 7 ASP A 240 ALA A 245 1 6
HELIX 8 8 THR A 308 ASP A 323 1 16
HELIX 9 9 THR A 331 LEU A 342 1 12
HELIX 10 10 THR A 347 ASP A 359 1 13
HELIX 11 11 PHE A 368 ARG A 377 1 10
HELIX 12 12 LYS A 380 ASP A 396 1 17
HELIX 13 13 SER A 404 GLY A 416 1 13
HELIX 14 14 THR A 420 ASP A 432 1 13
HELIX 15 15 TYR A 441 THR A 449 1 9
SHEET 1 A12 VAL A 63 ASP A 68 0
SHEET 2 A12 GLY A 73 ASN A 83 -1 O GLY A 73 N ASP A 68
SHEET 3 A12 VAL A 89 PRO A 100 -1 O GLN A 90 N HIS A 82
SHEET 4 A12 TYR A 250 PHE A 258 -1 O ILE A 251 N THR A 99
SHEET 5 A12 ASN A 263 GLU A 273 -1 O VAL A 270 N TYR A 250
SHEET 6 A12 THR A 276 ILE A 286 -1 O VAL A 278 N LYS A 271
SHEET 7 A12 VAL A 170 VAL A 180 1 N GLU A 175 O ILE A 281
SHEET 8 A12 HIS A 183 ASP A 194 -1 O PHE A 185 N GLY A 178
SHEET 9 A12 LYS A 199 CYS A 206 -1 O LYS A 199 N ASP A 194
SHEET 10 A12 HIS A 130 ALA A 140 -1 N MET A 131 O PHE A 204
SHEET 11 A12 HIS A 112 SER A 121 -1 N LYS A 119 O LEU A 134
SHEET 12 A12 VAL A 63 ASP A 68 -1 N ILE A 65 O HIS A 112
SHEET 1 B 2 THR A 329 ILE A 330 0
SHEET 2 B 2 ILE A 366 ASP A 367 -1 O ILE A 366 N ILE A 330
SHEET 1 C 2 TYR A 402 ILE A 403 0
SHEET 2 C 2 VAL A 439 ASN A 440 -1 O VAL A 439 N ILE A 403
LINK C LEU A 222 N1 CRO A 224 1555 1555 1.35
LINK C3 CRO A 224 N VAL A 226 1555 1555 1.34
LINK OD1 ASP A 323 CA CA A 452 1555 1555 2.27
LINK OD1 ASP A 325 CA CA A 452 1555 1555 2.48
LINK OD1 ASP A 327 CA CA A 452 1555 1555 2.46
LINK O THR A 329 CA CA A 452 1555 1555 2.12
LINK OE1 GLU A 334 CA CA A 452 1555 1555 2.55
LINK OE2 GLU A 334 CA CA A 452 1555 1555 2.48
LINK OD2 ASP A 359 CA CA A 453 1555 1555 2.19
LINK OD1 ASP A 361 CA CA A 453 1555 1555 2.36
LINK OD1 ASN A 363 CA CA A 453 1555 1555 2.34
LINK O THR A 365 CA CA A 453 1555 1555 2.45
LINK OE1 GLU A 370 CA CA A 453 1555 1555 2.46
LINK OE2 GLU A 370 CA CA A 453 1555 1555 2.64
LINK OD2 ASP A 396 CA CA A 454 1555 1555 2.34
LINK OD2 ASP A 398 CA CA A 454 1555 1555 2.44
LINK OD1 ASN A 400 CA CA A 454 1555 1555 2.43
LINK O TYR A 402 CA CA A 454 1555 1555 2.14
LINK OE1 GLU A 407 CA CA A 454 1555 1555 2.65
LINK OE2 GLU A 407 CA CA A 454 1555 1555 2.40
LINK OD1 ASP A 432 CA CA A 455 1555 1555 2.29
LINK OD2 ASP A 434 CA CA A 455 1555 1555 2.43
LINK OD1 ASP A 436 CA CA A 455 1555 1555 2.35
LINK O GLN A 438 CA CA A 455 1555 1555 2.35
LINK OE1 GLU A 443 CA CA A 455 1555 1555 2.52
LINK OE2 GLU A 443 CA CA A 455 1555 1555 2.69
LINK CA CA A 452 O HOH A 507 1555 1555 2.44
LINK CA CA A 453 O HOH A 518 1555 1555 2.45
LINK CA CA A 454 O HOH A 550 1555 1555 2.47
LINK CA CA A 455 O HOH A 525 1555 1555 2.38
CISPEP 1 MET A 246 PRO A 247 0 2.62
SITE 1 AC1 6 ASP A 323 ASP A 325 ASP A 327 THR A 329
SITE 2 AC1 6 GLU A 334 HOH A 507
SITE 1 AC2 6 ASP A 359 ASP A 361 ASN A 363 THR A 365
SITE 2 AC2 6 GLU A 370 HOH A 518
SITE 1 AC3 6 ASP A 396 ASP A 398 ASN A 400 TYR A 402
SITE 2 AC3 6 GLU A 407 HOH A 550
SITE 1 AC4 6 ASP A 432 ASP A 434 ASP A 436 GLN A 438
SITE 2 AC4 6 GLU A 443 HOH A 525
CRYST1 126.131 47.310 68.940 90.00 100.48 90.00 C 1 2 1 4
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 0.007928 0.000000 0.001467 0.00000
SCALE2 0.000000 0.021137 0.000000 0.00000
SCALE3 0.000000 0.000000 0.014751 0.00000
(ATOM LINES ARE NOT SHOWN.)
END
