HEADER FLUORESCENT PROTEIN 14-NOV-22 8HGI TITLE CRYSTAL STRUCTURE OF VNAR AGFP14 IN COMPLEX WITH GFP COMPND MOL_ID: 1; COMPND 2 MOLECULE: GFP; COMPND 3 CHAIN: A, B; COMPND 4 ENGINEERED: YES; COMPND 5 OTHER_DETAILS: SIMILAR TO WTGFP, THE CHROMOPHORE, CRQ, IS FORMED BY COMPND 6 THE RESIDUES OF GLN66-TYR67-GLY68.; COMPND 7 MOL_ID: 2; COMPND 8 MOLECULE: VNAR AGFP14; COMPND 9 CHAIN: C, D; COMPND 10 ENGINEERED: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: SYNTHETIC CONSTRUCT; SOURCE 3 ORGANISM_TAXID: 32630; SOURCE 4 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 5 EXPRESSION_SYSTEM_TAXID: 562; SOURCE 6 MOL_ID: 2; SOURCE 7 ORGANISM_SCIENTIFIC: CHILOSCYLLIUM PLAGIOSUM; SOURCE 8 ORGANISM_TAXID: 36176; SOURCE 9 EXPRESSION_SYSTEM: HOMO SAPIENS; SOURCE 10 EXPRESSION_SYSTEM_TAXID: 9606 KEYWDS GFP, AGFP14, NANOBODY, CHROMOPHORE, FLUORESCENT PROTEIN EXPDTA X-RAY DIFFRACTION AUTHOR P.ZHENG,C.ZHU,T.JIN REVDAT 4 20-NOV-24 8HGI 1 REMARK REVDAT 3 02-OCT-24 8HGI 1 COMPND SOURCE REMARK DBREF REVDAT 3 2 1 SEQRES HELIX SHEET LINK REVDAT 3 3 1 SCALE ATOM REVDAT 2 15-NOV-23 8HGI 1 LINK ATOM REVDAT 1 27-SEP-23 8HGI 0 JRNL AUTH Y.L.CHEN,X.X.XIE,P.ZHENG,C.ZHU,H.MA,Z.KHALID,Y.J.XIE, JRNL AUTH 2 Y.Z.DANG,Y.YE,N.SHENG,N.ZHONG,W.H.LEI,C.ZHANG,L.J.ZHANG, JRNL AUTH 3 T.JIN,M.J.CAO JRNL TITL SELECTION, IDENTIFICATION AND CRYSTAL STRUCTURE OF JRNL TITL 2 SHARK-DERIVED SINGLE-DOMAIN ANTIBODIES AGAINST A GREEN JRNL TITL 3 FLUORESCENT PROTEIN. JRNL REF INT.J.BIOL.MACROMOL. V. 247 25852 2023 JRNL REFN ISSN 0141-8130 JRNL PMID 37460076 JRNL DOI 10.1016/J.IJBIOMAC.2023.125852 REMARK 2 REMARK 2 RESOLUTION. 1.95 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : REFMAC 5.8.0267 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) : 1.95 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 59.71 REMARK 3 DATA CUTOFF (SIGMA(F)) : NULL REMARK 3 COMPLETENESS FOR RANGE (%) : 99.3 REMARK 3 NUMBER OF REFLECTIONS : 49389 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) : 0.189 REMARK 3 R VALUE (WORKING SET) : 0.186 REMARK 3 FREE R VALUE : 0.242 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 4.900 REMARK 3 FREE R VALUE TEST SET COUNT : 2536 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) : 1.95 REMARK 3 BIN RESOLUTION RANGE LOW (A) : 2.00 REMARK 3 REFLECTION IN BIN (WORKING SET) : 3426 REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 92.46 REMARK 3 BIN R VALUE (WORKING SET) : 0.2720 REMARK 3 BIN FREE R VALUE SET COUNT : 131 REMARK 3 BIN FREE R VALUE : 0.3170 REMARK 3 REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT. REMARK 3 PROTEIN ATOMS : 5373 REMARK 3 NUCLEIC ACID ATOMS : 0 REMARK 3 HETEROGEN ATOMS : 0 REMARK 3 SOLVENT ATOMS : 470 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : 27.60 REMARK 3 MEAN B VALUE (OVERALL, A**2) : 31.79 REMARK 3 OVERALL ANISOTROPIC B VALUE. REMARK 3 B11 (A**2) : 1.89000 REMARK 3 B22 (A**2) : 2.43000 REMARK 3 B33 (A**2) : -2.48000 REMARK 3 B12 (A**2) : 0.00000 REMARK 3 B13 (A**2) : -0.63000 REMARK 3 B23 (A**2) : 0.00000 REMARK 3 REMARK 3 ESTIMATED OVERALL COORDINATE ERROR. REMARK 3 ESU BASED ON R VALUE (A): 0.175 REMARK 3 ESU BASED ON FREE R VALUE (A): 0.165 REMARK 3 ESU BASED ON MAXIMUM LIKELIHOOD (A): 0.131 REMARK 3 ESU FOR B VALUES BASED ON MAXIMUM LIKELIHOOD (A**2): 4.730 REMARK 3 REMARK 3 CORRELATION COEFFICIENTS. REMARK 3 CORRELATION COEFFICIENT FO-FC : 0.960 REMARK 3 CORRELATION COEFFICIENT FO-FC FREE : 0.933 REMARK 3 REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES COUNT RMS WEIGHT REMARK 3 BOND LENGTHS REFINED ATOMS (A): 5506 ; 0.008 ; 0.013 REMARK 3 BOND LENGTHS OTHERS (A): 5077 ; 0.002 ; 0.017 REMARK 3 BOND ANGLES REFINED ATOMS (DEGREES): 7431 ; 1.514 ; 1.663 REMARK 3 BOND ANGLES OTHERS (DEGREES): 11765 ; 1.319 ; 1.589 REMARK 3 TORSION ANGLES, PERIOD 1 (DEGREES): 659 ; 6.938 ; 5.000 REMARK 3 TORSION ANGLES, PERIOD 2 (DEGREES): 287 ;31.185 ;21.777 REMARK 3 TORSION ANGLES, PERIOD 3 (DEGREES): 964 ;18.171 ;15.000 REMARK 3 TORSION ANGLES, PERIOD 4 (DEGREES): 33 ;16.281 ;15.000 REMARK 3 CHIRAL-CENTER RESTRAINTS (A**3): 701 ; 0.068 ; 0.200 REMARK 3 GENERAL PLANES REFINED ATOMS (A): 6143 ; 0.008 ; 0.020 REMARK 3 GENERAL PLANES OTHERS (A): 1259 ; 0.001 ; 0.020 REMARK 3 NON-BONDED CONTACTS REFINED ATOMS (A): NULL ; NULL ; NULL REMARK 3 NON-BONDED CONTACTS OTHERS (A): NULL ; NULL ; NULL REMARK 3 NON-BONDED TORSION REFINED ATOMS (A): NULL ; NULL ; NULL REMARK 3 NON-BONDED TORSION OTHERS (A): NULL ; NULL ; NULL REMARK 3 H-BOND (X...Y) REFINED ATOMS (A): NULL ; NULL ; NULL REMARK 3 H-BOND (X...Y) OTHERS (A): NULL ; NULL ; NULL REMARK 3 POTENTIAL METAL-ION REFINED ATOMS (A): NULL ; NULL ; NULL REMARK 3 POTENTIAL METAL-ION OTHERS (A): NULL ; NULL ; NULL REMARK 3 SYMMETRY VDW REFINED ATOMS (A): NULL ; NULL ; NULL REMARK 3 SYMMETRY VDW OTHERS (A): NULL ; NULL ; NULL REMARK 3 SYMMETRY H-BOND REFINED ATOMS (A): NULL ; NULL ; NULL 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): 2656 ; 2.512 ; 3.076 REMARK 3 MAIN-CHAIN BOND OTHER ATOMS (A**2): 2655 ; 2.511 ; 3.075 REMARK 3 MAIN-CHAIN ANGLE REFINED ATOMS (A**2): 3307 ; 3.644 ; 4.598 REMARK 3 MAIN-CHAIN ANGLE OTHER ATOMS (A**2): 3308 ; 3.644 ; 4.599 REMARK 3 SIDE-CHAIN BOND REFINED ATOMS (A**2): 2850 ; 3.498 ; 3.456 REMARK 3 SIDE-CHAIN BOND OTHER ATOMS (A**2): 2850 ; 3.498 ; 3.456 REMARK 3 SIDE-CHAIN ANGLE REFINED ATOMS (A**2): NULL ; NULL ; NULL REMARK 3 SIDE-CHAIN ANGLE OTHER ATOMS (A**2): 4125 ; 5.545 ; 5.000 REMARK 3 LONG RANGE B REFINED ATOMS (A**2): 5891 ; 7.267 ;34.629 REMARK 3 LONG RANGE B OTHER ATOMS (A**2): 5832 ; 7.260 ;34.431 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 8HGI COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY PDBJ ON 17-NOV-22. REMARK 100 THE DEPOSITION ID IS D_1300033553. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 26-AUG-22 REMARK 200 TEMPERATURE (KELVIN) : 100 REMARK 200 PH : 6.8 REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : Y REMARK 200 RADIATION SOURCE : SSRF REMARK 200 BEAMLINE : BL10U2 REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 0.97902 REMARK 200 MONOCHROMATOR : NULL REMARK 200 OPTICS : NULL REMARK 200 REMARK 200 DETECTOR TYPE : PIXEL REMARK 200 DETECTOR MANUFACTURER : DECTRIS EIGER X 9M REMARK 200 INTENSITY-INTEGRATION SOFTWARE : DIALS 0.7.7 REMARK 200 DATA SCALING SOFTWARE : SCALEPACK 0.5.28 REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 51988 REMARK 200 RESOLUTION RANGE HIGH (A) : 1.950 REMARK 200 RESOLUTION RANGE LOW (A) : 102.800 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : NULL REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 99.4 REMARK 200 DATA REDUNDANCY : 5.800 REMARK 200 R MERGE (I) : 0.07200 REMARK 200 R SYM (I) : NULL REMARK 200 FOR THE DATA SET : 10.3000 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 1.95 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 2.00 REMARK 200 COMPLETENESS FOR SHELL (%) : 92.7 REMARK 200 DATA REDUNDANCY IN SHELL : 3.80 REMARK 200 R MERGE FOR SHELL (I) : 0.52300 REMARK 200 R SYM FOR SHELL (I) : NULL REMARK 200 FOR SHELL : 2.500 REMARK 200 REMARK 200 DIFFRACTION PROTOCOL: SINGLE WAVELENGTH REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: MOLECULAR REPLACEMENT REMARK 200 SOFTWARE USED: PHASER 2.8.3 REMARK 200 STARTING MODEL: 4PPK, 7FBJ REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 43.83 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.19 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 18%(W/V) PEG8000 0.1M NA/K PO4 PH6.8, REMARK 280 VAPOR DIFFUSION, SITTING DROP, TEMPERATURE 291K REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 1 21 1 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 -X,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 51.39850 REMARK 290 SMTRY3 2 0.000000 0.000000 -1.000000 0.00000 REMARK 290 REMARK 290 REMARK: NULL REMARK 300 REMARK 300 BIOMOLECULE: 1, 2 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: A, C 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 REMARK 350 BIOMOLECULE: 2 REMARK 350 AUTHOR DETERMINED BIOLOGICAL UNIT: DIMERIC REMARK 350 APPLY THE FOLLOWING TO CHAINS: B, D 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. (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 GLY A 2 REMARK 465 ALA A 3 REMARK 465 HIS A 4 REMARK 465 ALA A 228 REMARK 465 LYS A 229 REMARK 465 ALA A 230 REMARK 465 SER A 231 REMARK 465 GLY A 232 REMARK 465 LYS A 233 REMARK 465 PRO A 234 REMARK 465 ILE A 235 REMARK 465 PRO A 236 REMARK 465 ASN A 237 REMARK 465 PRO A 238 REMARK 465 LEU A 239 REMARK 465 LEU A 240 REMARK 465 GLY A 241 REMARK 465 LEU A 242 REMARK 465 ASP A 243 REMARK 465 SER A 244 REMARK 465 THR A 245 REMARK 465 MET B 1 REMARK 465 GLY B 2 REMARK 465 ALA B 3 REMARK 465 HIS B 4 REMARK 465 LEU B 224 REMARK 465 PRO B 225 REMARK 465 SER B 226 REMARK 465 GLN B 227 REMARK 465 ALA B 228 REMARK 465 LYS B 229 REMARK 465 ALA B 230 REMARK 465 SER B 231 REMARK 465 GLY B 232 REMARK 465 LYS B 233 REMARK 465 PRO B 234 REMARK 465 ILE B 235 REMARK 465 PRO B 236 REMARK 465 ASN B 237 REMARK 465 PRO B 238 REMARK 465 LEU B 239 REMARK 465 LEU B 240 REMARK 465 GLY B 241 REMARK 465 LEU B 242 REMARK 465 ASP B 243 REMARK 465 SER B 244 REMARK 465 THR B 245 REMARK 465 GLY C 116 REMARK 465 SER C 117 REMARK 465 GLU C 118 REMARK 465 ASN C 119 REMARK 465 LEU C 120 REMARK 465 TYR C 121 REMARK 465 PHE C 122 REMARK 465 GLN C 123 REMARK 465 ARG D 115 REMARK 465 GLY D 116 REMARK 465 SER D 117 REMARK 465 GLU D 118 REMARK 465 ASN D 119 REMARK 465 LEU D 120 REMARK 465 TYR D 121 REMARK 465 PHE D 122 REMARK 465 GLN D 123 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 ASP C 33 -113.91 -110.18 REMARK 500 ASP D 33 -100.02 -104.77 REMARK 500 REMARK 500 REMARK: NULL DBREF 8HGI A 1 245 PDB 8HGI 8HGI 1 245 DBREF 8HGI B 1 245 PDB 8HGI 8HGI 1 245 DBREF 8HGI C 1 123 PDB 8HGI 8HGI 1 123 DBREF 8HGI D 1 123 PDB 8HGI 8HGI 1 123 SEQRES 1 A 243 MET GLY ALA HIS ALA SER VAL ILE LYS PRO GLU MET LYS SEQRES 2 A 243 ILE LYS LEU ARG MET GLU GLY ALA VAL ASN GLY HIS LYS SEQRES 3 A 243 PHE VAL ILE GLU GLY GLU GLY ILE GLY LYS PRO TYR GLU SEQRES 4 A 243 GLY THR GLN THR LEU ASP LEU THR VAL LYS GLU GLY ALA SEQRES 5 A 243 PRO LEU PRO PHE SER TYR ASP ILE LEU THR PRO ALA PHE SEQRES 6 A 243 CRQ ASN ARG ALA PHE THR LYS TYR PRO LYS ASP ILE PRO SEQRES 7 A 243 ASP TYR PHE LYS GLN ALA PHE PRO GLU GLY TYR SER TRP SEQRES 8 A 243 GLU ARG SER MET THR TYR GLU ASP GLN GLY ILE CYS ILE SEQRES 9 A 243 ALA THR SER ASP ILE THR MET GLU GLY ASP CYS PHE PHE SEQRES 10 A 243 TYR LYS ILE ARG PHE ASP GLY THR ASN PHE PRO PRO ASN SEQRES 11 A 243 GLY PRO VAL MET GLN LYS LYS THR LEU LYS TRP GLU PRO SEQRES 12 A 243 SER THR GLU LYS MET TYR VAL ARG ASP GLY VAL LEU LYS SEQRES 13 A 243 GLY ASP VAL ASN MET ALA LEU LEU LEU GLU GLY GLY GLY SEQRES 14 A 243 HIS TYR ARG CYS ASP PHE LYS THR THR TYR LYS ALA LYS SEQRES 15 A 243 LYS ASP VAL ARG LEU PRO ASP ALA HIS LYS VAL ASP HIS SEQRES 16 A 243 ARG ILE GLU ILE LEU SER HIS ASP LYS ASP TYR ASN LYS SEQRES 17 A 243 VAL ARG LEU TYR GLU HIS ALA GLU ALA ARG TYR SER MET SEQRES 18 A 243 LEU PRO SER GLN ALA LYS ALA SER GLY LYS PRO ILE PRO SEQRES 19 A 243 ASN PRO LEU LEU GLY LEU ASP SER THR SEQRES 1 B 243 MET GLY ALA HIS ALA SER VAL ILE LYS PRO GLU MET LYS SEQRES 2 B 243 ILE LYS LEU ARG MET GLU GLY ALA VAL ASN GLY HIS LYS SEQRES 3 B 243 PHE VAL ILE GLU GLY GLU GLY ILE GLY LYS PRO TYR GLU SEQRES 4 B 243 GLY THR GLN THR LEU ASP LEU THR VAL LYS GLU GLY ALA SEQRES 5 B 243 PRO LEU PRO PHE SER TYR ASP ILE LEU THR PRO ALA PHE SEQRES 6 B 243 CRQ ASN ARG ALA PHE THR LYS TYR PRO LYS ASP ILE PRO SEQRES 7 B 243 ASP TYR PHE LYS GLN ALA PHE PRO GLU GLY TYR SER TRP SEQRES 8 B 243 GLU ARG SER MET THR TYR GLU ASP GLN GLY ILE CYS ILE SEQRES 9 B 243 ALA THR SER ASP ILE THR MET GLU GLY ASP CYS PHE PHE SEQRES 10 B 243 TYR LYS ILE ARG PHE ASP GLY THR ASN PHE PRO PRO ASN SEQRES 11 B 243 GLY PRO VAL MET GLN LYS LYS THR LEU LYS TRP GLU PRO SEQRES 12 B 243 SER THR GLU LYS MET TYR VAL ARG ASP GLY VAL LEU LYS SEQRES 13 B 243 GLY ASP VAL ASN MET ALA LEU LEU LEU GLU GLY GLY GLY SEQRES 14 B 243 HIS TYR ARG CYS ASP PHE LYS THR THR TYR LYS ALA LYS SEQRES 15 B 243 LYS ASP VAL ARG LEU PRO ASP ALA HIS LYS VAL ASP HIS SEQRES 16 B 243 ARG ILE GLU ILE LEU SER HIS ASP LYS ASP TYR ASN LYS SEQRES 17 B 243 VAL ARG LEU TYR GLU HIS ALA GLU ALA ARG TYR SER MET SEQRES 18 B 243 LEU PRO SER GLN ALA LYS ALA SER GLY LYS PRO ILE PRO SEQRES 19 B 243 ASN PRO LEU LEU GLY LEU ASP SER THR SEQRES 1 C 123 THR GLN ARG VAL GLU GLN THR PRO THR THR THR THR LYS SEQRES 2 C 123 GLU ALA GLY GLU SER LEU THR ILE ASN CYS VAL LEU ARG SEQRES 3 C 123 ASP SER SER CYS ALA LEU ASP SER THR TYR TRP TYR PHE SEQRES 4 C 123 THR LYS LYS GLY ALA THR LYS LYS GLU SER LEU SER ASN SEQRES 5 C 123 GLY GLY ARG TYR ALA GLU THR VAL ASN LYS ALA SER LYS SEQRES 6 C 123 SER PHE SER LEU ARG ILE SER ASP LEU ARG VAL GLU ASP SEQRES 7 C 123 SER GLY THR TYR HIS CYS LYS ALA TYR TYR SER TRP ASP SEQRES 8 C 123 ALA ASN CYS TRP ASN LEU ARG TYR SER TYR ILE GLU GLY SEQRES 9 C 123 GLY GLY THR ILE LEU THR VAL LYS PRO SER ARG GLY SER SEQRES 10 C 123 GLU ASN LEU TYR PHE GLN SEQRES 1 D 123 THR GLN ARG VAL GLU GLN THR PRO THR THR THR THR LYS SEQRES 2 D 123 GLU ALA GLY GLU SER LEU THR ILE ASN CYS VAL LEU ARG SEQRES 3 D 123 ASP SER SER CYS ALA LEU ASP SER THR TYR TRP TYR PHE SEQRES 4 D 123 THR LYS LYS GLY ALA THR LYS LYS GLU SER LEU SER ASN SEQRES 5 D 123 GLY GLY ARG TYR ALA GLU THR VAL ASN LYS ALA SER LYS SEQRES 6 D 123 SER PHE SER LEU ARG ILE SER ASP LEU ARG VAL GLU ASP SEQRES 7 D 123 SER GLY THR TYR HIS CYS LYS ALA TYR TYR SER TRP ASP SEQRES 8 D 123 ALA ASN CYS TRP ASN LEU ARG TYR SER TYR ILE GLU GLY SEQRES 9 D 123 GLY GLY THR ILE LEU THR VAL LYS PRO SER ARG GLY SER SEQRES 10 D 123 GLU ASN LEU TYR PHE GLN HET CRQ A 66 24 HET CRQ B 66 24 HETNAM CRQ [2-(3-CARBAMOYL-1-IMINO-PROPYL)-4-(4-HYDROXY- HETNAM 2 CRQ BENZYLIDENE)-5-OXO-4,5-DIHYDRO-IMIDAZOL-1-YL]-ACETIC HETNAM 3 CRQ ACID HETSYN CRQ CHROMOPHORE (GLN-TYR-GLY) FORMUL 1 CRQ 2(C16 H16 N4 O5) FORMUL 5 HOH *470(H2 O) HELIX 1 AA1 SER A 57 PHE A 65 5 9 HELIX 2 AA2 ASP A 81 ALA A 86 1 6 HELIX 3 AA3 SER B 57 PHE B 65 5 9 HELIX 4 AA4 ASP B 81 ALA B 86 1 6 HELIX 5 AA5 ARG C 75 SER C 79 5 5 HELIX 6 AA6 ASP C 91 ARG C 98 5 8 HELIX 7 AA7 ARG D 75 SER D 79 5 5 HELIX 8 AA8 ASP D 91 ARG D 98 5 8 SHEET 1 AA113 THR A 140 TRP A 143 0 SHEET 2 AA113 VAL A 156 LEU A 167 -1 O LEU A 166 N LEU A 141 SHEET 3 AA113 HIS A 172 ALA A 183 -1 O TYR A 181 N LEU A 157 SHEET 4 AA113 TYR A 91 TYR A 99 -1 N SER A 92 O LYS A 182 SHEET 5 AA113 ILE A 104 GLU A 114 -1 O ILE A 111 N TYR A 91 SHEET 6 AA113 CYS A 117 THR A 127 -1 O PHE A 119 N THR A 112 SHEET 7 AA113 MET A 12 VAL A 22 1 N ARG A 17 O ILE A 122 SHEET 8 AA113 HIS A 25 LYS A 36 -1 O ILE A 29 N MET A 18 SHEET 9 AA113 THR A 41 GLU A 50 -1 O ASP A 45 N GLU A 32 SHEET 10 AA113 LYS A 210 ARG A 220 -1 O VAL A 211 N LEU A 46 SHEET 11 AA113 HIS A 193 HIS A 204 -1 N LYS A 194 O ARG A 220 SHEET 12 AA113 SER A 146 ARG A 153 -1 N MET A 150 O HIS A 193 SHEET 13 AA113 VAL A 156 LEU A 167 -1 O LYS A 158 N TYR A 151 SHEET 1 AA213 THR B 140 TRP B 143 0 SHEET 2 AA213 VAL B 156 LEU B 167 -1 O LEU B 166 N LEU B 141 SHEET 3 AA213 HIS B 172 ALA B 183 -1 O THR B 179 N GLY B 159 SHEET 4 AA213 TYR B 91 TYR B 99 -1 N SER B 92 O LYS B 182 SHEET 5 AA213 ILE B 104 GLU B 114 -1 O ILE B 111 N TYR B 91 SHEET 6 AA213 CYS B 117 THR B 127 -1 O PHE B 119 N THR B 112 SHEET 7 AA213 MET B 12 VAL B 22 1 N ARG B 17 O ILE B 122 SHEET 8 AA213 HIS B 25 LYS B 36 -1 O ILE B 29 N MET B 18 SHEET 9 AA213 THR B 41 GLU B 50 -1 O ASP B 45 N GLU B 32 SHEET 10 AA213 LYS B 210 ARG B 220 -1 O VAL B 211 N LEU B 46 SHEET 11 AA213 HIS B 193 HIS B 204 -1 N LYS B 194 O ARG B 220 SHEET 12 AA213 SER B 146 ARG B 153 -1 N GLU B 148 O VAL B 195 SHEET 13 AA213 VAL B 156 LEU B 167 -1 O LYS B 158 N TYR B 151 SHEET 1 AA3 4 ARG C 3 THR C 7 0 SHEET 2 AA3 4 LEU C 19 ARG C 26 -1 O VAL C 24 N GLU C 5 SHEET 3 AA3 4 SER C 66 ILE C 71 -1 O ILE C 71 N LEU C 19 SHEET 4 AA3 4 TYR C 56 ASN C 61 -1 N ASN C 61 O SER C 66 SHEET 1 AA4 5 THR C 10 GLU C 14 0 SHEET 2 AA4 5 THR C 107 LYS C 112 1 O THR C 110 N LYS C 13 SHEET 3 AA4 5 GLY C 80 TYR C 88 -1 N GLY C 80 O LEU C 109 SHEET 4 AA4 5 LEU C 32 THR C 40 -1 N THR C 40 O THR C 81 SHEET 5 AA4 5 GLU C 48 SER C 49 -1 O GLU C 48 N PHE C 39 SHEET 1 AA5 4 THR C 10 GLU C 14 0 SHEET 2 AA5 4 THR C 107 LYS C 112 1 O THR C 110 N LYS C 13 SHEET 3 AA5 4 GLY C 80 TYR C 88 -1 N GLY C 80 O LEU C 109 SHEET 4 AA5 4 TYR C 101 GLU C 103 -1 O ILE C 102 N ALA C 86 SHEET 1 AA6 4 ARG D 3 THR D 7 0 SHEET 2 AA6 4 LEU D 19 ARG D 26 -1 O VAL D 24 N GLU D 5 SHEET 3 AA6 4 SER D 66 ILE D 71 -1 O LEU D 69 N ILE D 21 SHEET 4 AA6 4 TYR D 56 ASN D 61 -1 N ASN D 61 O SER D 66 SHEET 1 AA7 5 THR D 10 GLU D 14 0 SHEET 2 AA7 5 THR D 107 LYS D 112 1 O LYS D 112 N LYS D 13 SHEET 3 AA7 5 GLY D 80 TYR D 88 -1 N GLY D 80 O LEU D 109 SHEET 4 AA7 5 LEU D 32 THR D 40 -1 N ASP D 33 O TYR D 87 SHEET 5 AA7 5 GLU D 48 SER D 49 -1 O GLU D 48 N PHE D 39 SHEET 1 AA8 4 THR D 10 GLU D 14 0 SHEET 2 AA8 4 THR D 107 LYS D 112 1 O LYS D 112 N LYS D 13 SHEET 3 AA8 4 GLY D 80 TYR D 88 -1 N GLY D 80 O LEU D 109 SHEET 4 AA8 4 TYR D 101 GLU D 103 -1 O ILE D 102 N ALA D 86 SSBOND 1 CYS C 23 CYS C 84 1555 1555 2.07 SSBOND 2 CYS C 30 CYS C 94 1555 1555 2.07 SSBOND 3 CYS D 23 CYS D 84 1555 1555 2.09 SSBOND 4 CYS D 30 CYS D 94 1555 1555 2.09 LINK C PHE A 65 N1 CRQ A 66 1555 1555 1.43 LINK C3 CRQ A 66 N ASN A 69 1555 1555 1.47 LINK C PHE B 65 N1 CRQ B 66 1555 1555 1.43 LINK C3 CRQ B 66 N ASN B 69 1555 1555 1.59 CISPEP 1 ALA A 52 PRO A 53 0 -15.94 CISPEP 2 PHE A 87 PRO A 88 0 6.05 CISPEP 3 ALA B 52 PRO B 53 0 -9.43 CISPEP 4 PHE B 87 PRO B 88 0 10.26 CISPEP 5 THR C 7 PRO C 8 0 -4.44 CISPEP 6 THR D 7 PRO D 8 0 -4.26 CRYST1 59.449 102.797 66.620 90.00 116.33 90.00 P 1 21 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 0.016821 0.000000 0.008324 0.00000 SCALE2 0.000000 0.009728 0.000000 0.00000 SCALE3 0.000000 0.000000 0.016748 0.00000