HEADER BLOOD CLOTTING 23-FEB-21 7NMU TITLE CRYSTAL STRUCTURE OF HUMAN PLATELET GLYCOPROTEIN VI IN COMPLEX WITH AN TITLE 2 INHIBITORY NANOBODY. COMPND MOL_ID: 1; COMPND 2 MOLECULE: PLATELET GLYCOPROTEIN VI; COMPND 3 CHAIN: AAA, BBB; COMPND 4 SYNONYM: GPVI,GLYCOPROTEIN 6; COMPND 5 ENGINEERED: YES; COMPND 6 MUTATION: YES; COMPND 7 MOL_ID: 2; COMPND 8 MOLECULE: NANOBODY 2; COMPND 9 CHAIN: DDD, CCC; 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: GP6; SOURCE 6 EXPRESSION_SYSTEM: HOMO SAPIENS; SOURCE 7 EXPRESSION_SYSTEM_TAXID: 9606; SOURCE 8 MOL_ID: 2; SOURCE 9 ORGANISM_SCIENTIFIC: LAMA GLAMA; SOURCE 10 ORGANISM_TAXID: 9844; SOURCE 11 EXPRESSION_SYSTEM: ESCHERICHIA COLI K-12; SOURCE 12 EXPRESSION_SYSTEM_TAXID: 83333; SOURCE 13 EXPRESSION_SYSTEM_VARIANT: WK6 KEYWDS NANOBODY, COMPLEX, DOMAIN-SWAP, BLOOD CLOTTING EXPDTA X-RAY DIFFRACTION AUTHOR A.SLATER,E.JONAS,S.P.WATSON JRNL AUTH A.SLATER,Y.DI,J.C.CLARK,N.J.JOOSS,E.M.MARTIN,F.ALENAZY, JRNL AUTH 2 M.R.THOMAS,R.A.S.ARIENS,A.B.HERR,N.S.POULTER,J.EMSLEY, JRNL AUTH 3 S.P.WATSON JRNL TITL STRUCTURAL CHARACTERIZATION OF A NOVEL GPVI-NANOBODY COMPLEX JRNL TITL 2 REVEALS A BIOLOGICALLY ACTIVE DOMAIN-SWAPPED GPVI DIMER. JRNL REF BLOOD V. 137 3443 2021 JRNL REFN ESSN 1528-0020 JRNL PMID 33512486 JRNL DOI 10.1182/BLOOD.2020009440 REMARK 2 REMARK 2 RESOLUTION. 2.50 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : REFMAC 5.8.0257 REMARK 3 AUTHORS : MURSHUDOV,SKUBAK,LEBEDEV,PANNU,STEINER, REMARK 3 : NICHOLLS,WINN,LONG,VAGIN REMARK 3 REMARK 3 REFINEMENT TARGET : NULL REMARK 3 REMARK 3 DATA USED IN REFINEMENT. REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 2.50 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 70.08 REMARK 3 DATA CUTOFF (SIGMA(F)) : NULL REMARK 3 COMPLETENESS FOR RANGE (%) : 99.9 REMARK 3 NUMBER OF REFLECTIONS : 26173 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 CROSS-VALIDATION METHOD : FREE R-VALUE REMARK 3 FREE R VALUE TEST SET SELECTION : NULL REMARK 3 R VALUE (WORKING + TEST SET) : NULL REMARK 3 R VALUE (WORKING SET) : 0.174 REMARK 3 FREE R VALUE : 0.235 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 5.078 REMARK 3 FREE R VALUE TEST SET COUNT : 1329 REMARK 3 REMARK 3 FIT IN THE HIGHEST RESOLUTION BIN. REMARK 3 TOTAL NUMBER OF BINS USED : NULL REMARK 3 BIN RESOLUTION RANGE HIGH (A) : 2.50 REMARK 3 BIN RESOLUTION RANGE LOW (A) : 2.56 REMARK 3 REFLECTION IN BIN (WORKING SET) : 1761 REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 98.99 REMARK 3 BIN R VALUE (WORKING SET) : 0.2200 REMARK 3 BIN FREE R VALUE SET COUNT : 93 REMARK 3 BIN FREE R VALUE : 0.2850 REMARK 3 REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT. REMARK 3 PROTEIN ATOMS : 4664 REMARK 3 NUCLEIC ACID ATOMS : 0 REMARK 3 HETEROGEN ATOMS : 1 REMARK 3 SOLVENT ATOMS : 269 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : NULL REMARK 3 MEAN B VALUE (OVERALL, A**2) : 40.54 REMARK 3 OVERALL ANISOTROPIC B VALUE. REMARK 3 B11 (A**2) : 0.81800 REMARK 3 B22 (A**2) : -0.83900 REMARK 3 B33 (A**2) : 0.02000 REMARK 3 B12 (A**2) : 0.00000 REMARK 3 B13 (A**2) : 0.00000 REMARK 3 B23 (A**2) : 0.00000 REMARK 3 REMARK 3 ESTIMATED OVERALL COORDINATE ERROR. REMARK 3 ESU BASED ON R VALUE (A): 0.435 REMARK 3 ESU BASED ON FREE R VALUE (A): 0.266 REMARK 3 ESU BASED ON MAXIMUM LIKELIHOOD (A): 0.184 REMARK 3 ESU FOR B VALUES BASED ON MAXIMUM LIKELIHOOD (A**2): 8.315 REMARK 3 REMARK 3 CORRELATION COEFFICIENTS. REMARK 3 CORRELATION COEFFICIENT FO-FC : 0.953 REMARK 3 CORRELATION COEFFICIENT FO-FC FREE : 0.914 REMARK 3 REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES COUNT RMS WEIGHT REMARK 3 BOND LENGTHS REFINED ATOMS (A): 4792 ; 0.008 ; 0.013 REMARK 3 BOND LENGTHS OTHERS (A): 4272 ; 0.001 ; 0.017 REMARK 3 BOND ANGLES REFINED ATOMS (DEGREES): 6532 ; 1.551 ; 1.652 REMARK 3 BOND ANGLES OTHERS (DEGREES): 9964 ; 1.211 ; 1.570 REMARK 3 TORSION ANGLES, PERIOD 1 (DEGREES): 603 ; 8.728 ; 5.000 REMARK 3 TORSION ANGLES, PERIOD 2 (DEGREES): 226 ;33.892 ;21.947 REMARK 3 TORSION ANGLES, PERIOD 3 (DEGREES): 729 ;16.113 ;15.000 REMARK 3 TORSION ANGLES, PERIOD 4 (DEGREES): 28 ;18.373 ;15.000 REMARK 3 CHIRAL-CENTER RESTRAINTS (A**3): 611 ; 0.065 ; 0.200 REMARK 3 GENERAL PLANES REFINED ATOMS (A): 5389 ; 0.006 ; 0.020 REMARK 3 GENERAL PLANES OTHERS (A): 1007 ; 0.001 ; 0.020 REMARK 3 NON-BONDED CONTACTS REFINED ATOMS (A): 712 ; 0.186 ; 0.200 REMARK 3 NON-BONDED CONTACTS OTHERS (A): 48 ; 0.172 ; 0.200 REMARK 3 NON-BONDED TORSION REFINED ATOMS (A): 2220 ; 0.168 ; 0.200 REMARK 3 NON-BONDED TORSION OTHERS (A): NULL ; NULL ; NULL REMARK 3 H-BOND (X...Y) REFINED ATOMS (A): 235 ; 0.169 ; 0.200 REMARK 3 H-BOND (X...Y) OTHERS (A): NULL ; NULL ; NULL REMARK 3 POTENTIAL METAL-ION REFINED ATOMS (A): 3 ; 0.197 ; 0.200 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): 1 ; 0.028 ; 0.200 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): 2430 ; 3.368 ; 4.135 REMARK 3 MAIN-CHAIN BOND OTHER ATOMS (A**2): 2429 ; 3.369 ; 4.134 REMARK 3 MAIN-CHAIN ANGLE REFINED ATOMS (A**2): 3027 ; 5.236 ; 6.178 REMARK 3 MAIN-CHAIN ANGLE OTHER ATOMS (A**2): 3028 ; 5.235 ; 6.180 REMARK 3 SIDE-CHAIN BOND REFINED ATOMS (A**2): 2362 ; 3.784 ; 4.576 REMARK 3 SIDE-CHAIN BOND OTHER ATOMS (A**2): 2363 ; 3.783 ; 4.578 REMARK 3 SIDE-CHAIN ANGLE REFINED ATOMS (A**2): 3505 ; 5.954 ; 6.698 REMARK 3 SIDE-CHAIN ANGLE OTHER ATOMS (A**2): 3506 ; 5.953 ; 6.700 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 BULK SOLVENT 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 THEIR REMARK 3 RIDING POSITIONS REMARK 4 REMARK 4 7NMU COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY PDBE. REMARK 100 THE DEPOSITION ID IS D_1292114113. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 06-OCT-19 REMARK 200 TEMPERATURE (KELVIN) : 283.15 REMARK 200 PH : NULL REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : Y REMARK 200 RADIATION SOURCE : DIAMOND REMARK 200 BEAMLINE : I24 REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 0.96864 REMARK 200 MONOCHROMATOR : NULL REMARK 200 OPTICS : NULL REMARK 200 REMARK 200 DETECTOR TYPE : PIXEL REMARK 200 DETECTOR MANUFACTURER : DECTRIS PILATUS3 6M REMARK 200 INTENSITY-INTEGRATION SOFTWARE : XIA2 REMARK 200 DATA SCALING SOFTWARE : XIA2 REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 26173 REMARK 200 RESOLUTION RANGE HIGH (A) : 2.498 REMARK 200 RESOLUTION RANGE LOW (A) : 84.750 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : NULL REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 98.6 REMARK 200 DATA REDUNDANCY : 6.300 REMARK 200 R MERGE (I) : 0.12200 REMARK 200 R SYM (I) : NULL REMARK 200 FOR THE DATA SET : 8.6700 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 2.50 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 2.56 REMARK 200 COMPLETENESS FOR SHELL (%) : NULL REMARK 200 DATA REDUNDANCY IN SHELL : NULL REMARK 200 R MERGE FOR SHELL (I) : NULL REMARK 200 R SYM FOR SHELL (I) : NULL REMARK 200 FOR SHELL : NULL 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: 2GI7 REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 52.49 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.59 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 0.2 M CALCIUM ACETATE, 0.1M SODIUM REMARK 280 CACODYLATE PH 6.5, 18% PEG8K, VAPOR DIFFUSION, SITTING DROP, REMARK 280 TEMPERATURE 283.15K REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 21 21 21 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 -X+1/2,-Y,Z+1/2 REMARK 290 3555 -X,Y+1/2,-Z+1/2 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 34.95300 REMARK 290 SMTRY2 2 0.000000 -1.000000 0.000000 0.00000 REMARK 290 SMTRY3 2 0.000000 0.000000 1.000000 62.02050 REMARK 290 SMTRY1 3 -1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 3 0.000000 1.000000 0.000000 42.37550 REMARK 290 SMTRY3 3 0.000000 0.000000 -1.000000 62.02050 REMARK 290 SMTRY1 4 1.000000 0.000000 0.000000 34.95300 REMARK 290 SMTRY2 4 0.000000 -1.000000 0.000000 42.37550 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: TETRAMERIC REMARK 350 APPLY THE FOLLOWING TO CHAINS: AAA, BBB, DDD, CCC 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 ASP A 1 REMARK 465 LYS A 2 REMARK 465 LEU A 3 REMARK 465 ALA A 4 REMARK 465 SER A 5 REMARK 465 SER A 6 REMARK 465 GLY A 7 REMARK 465 PRO A 8 REMARK 465 LYS A 139 REMARK 465 ASN A 140 REMARK 465 PRO A 141 REMARK 465 GLU A 142 REMARK 465 ARG A 143 REMARK 465 TRP A 144 REMARK 465 TYR A 145 REMARK 465 ARG A 146 REMARK 465 ILE A 191 REMARK 465 GLU A 192 REMARK 465 GLY A 193 REMARK 465 ARG A 194 REMARK 465 ASP B 1 REMARK 465 LYS B 2 REMARK 465 LEU B 3 REMARK 465 ALA B 4 REMARK 465 SER B 5 REMARK 465 LYS B 139 REMARK 465 ASN B 140 REMARK 465 PRO B 141 REMARK 465 GLU B 142 REMARK 465 ARG B 143 REMARK 465 TRP B 144 REMARK 465 TYR B 145 REMARK 465 ARG B 146 REMARK 465 GLU B 192 REMARK 465 GLY B 193 REMARK 465 ARG B 194 REMARK 465 ARG D 130 REMARK 465 PRO C 129 REMARK 465 ARG C 130 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 H THRAA 155 HD1 HISAA 158 1.23 REMARK 500 H THRBB 155 HD1 HISBB 158 1.32 REMARK 500 HH21 ARGAA 50 HG SERCC 99 1.32 REMARK 500 OG1 THRBB 161 OE1 GLUBB 183 2.16 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 GLUAA 25 9.46 87.84 REMARK 500 VALAA 38 139.07 -21.35 REMARK 500 ASPAA 39 -50.45 -123.79 REMARK 500 THRAA 153 72.70 50.20 REMARK 500 ASPBB 53 50.94 -100.44 REMARK 500 THRDD 57 125.58 -32.28 REMARK 500 LEUCC 127 32.91 -83.42 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 201 CA REMARK 620 N RES CSSEQI ATOM REMARK 620 1 GLU D 6 OE1 REMARK 620 2 GLY D 119 O 89.0 REMARK 620 3 HOH D 219 O 73.3 113.1 REMARK 620 4 GLU C 6 OE1 70.6 113.5 2.9 REMARK 620 5 GLY C 119 O 71.5 115.6 3.4 2.2 REMARK 620 6 HOH C 310 O 72.1 113.5 1.3 1.6 2.3 REMARK 620 N 1 2 3 4 5 DBREF 7NMUAA 6 188 UNP Q9HCN6 GPVI_HUMAN 22 204 DBREF 7NMUBB 6 188 UNP Q9HCN6 GPVI_HUMAN 22 204 DBREF 7NMUDD 1 130 PDB 7NMU 7NMU 1 130 DBREF 7NMUCC 1 130 PDB 7NMU 7NMU 1 130 SEQADV 7NMU ASPAA 1 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU LYSAA 2 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU LEUAA 3 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU ALAAA 4 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU SERAA 5 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU GLNAA 76 UNP Q9HCN6 ASN 92 ENGINEERED MUTATION SEQADV 7NMU ASPAA 189 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU PROAA 190 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU ILEAA 191 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU GLUAA 192 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU GLYAA 193 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU ARGAA 194 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU ASPBB 1 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU LYSBB 2 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU LEUBB 3 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU ALABB 4 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU SERBB 5 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU GLNBB 76 UNP Q9HCN6 ASN 92 ENGINEERED MUTATION SEQADV 7NMU ASPBB 189 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU PROBB 190 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU ILEBB 191 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU GLUBB 192 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU GLYBB 193 UNP Q9HCN6 EXPRESSION TAG SEQADV 7NMU ARGBB 194 UNP Q9HCN6 EXPRESSION TAG SEQRES 1AA 194 ASP LYS LEU ALA SER SER GLY PRO LEU PRO LYS PRO SER SEQRES 2AA 194 LEU GLN ALA LEU PRO SER SER LEU VAL PRO LEU GLU LYS SEQRES 3AA 194 PRO VAL THR LEU ARG CYS GLN GLY PRO PRO GLY VAL ASP SEQRES 4AA 194 LEU TYR ARG LEU GLU LYS LEU SER SER SER ARG TYR GLN SEQRES 5AA 194 ASP GLN ALA VAL LEU PHE ILE PRO ALA MET LYS ARG SER SEQRES 6AA 194 LEU ALA GLY ARG TYR ARG CYS SER TYR GLN GLN GLY SER SEQRES 7AA 194 LEU TRP SER LEU PRO SER ASP GLN LEU GLU LEU VAL ALA SEQRES 8AA 194 THR GLY VAL PHE ALA LYS PRO SER LEU SER ALA GLN PRO SEQRES 9AA 194 GLY PRO ALA VAL SER SER GLY GLY ASP VAL THR LEU GLN SEQRES 10AA 194 CYS GLN THR ARG TYR GLY PHE ASP GLN PHE ALA LEU TYR SEQRES 11AA 194 LYS GLU GLY ASP PRO ALA PRO TYR LYS ASN PRO GLU ARG SEQRES 12AA 194 TRP TYR ARG ALA SER PHE PRO ILE ILE THR VAL THR ALA SEQRES 13AA 194 ALA HIS SER GLY THR TYR ARG CYS TYR SER PHE SER SER SEQRES 14AA 194 ARG ASP PRO TYR LEU TRP SER ALA PRO SER ASP PRO LEU SEQRES 15AA 194 GLU LEU VAL VAL THR GLY ASP PRO ILE GLU GLY ARG SEQRES 1BB 194 ASP LYS LEU ALA SER SER GLY PRO LEU PRO LYS PRO SER SEQRES 2BB 194 LEU GLN ALA LEU PRO SER SER LEU VAL PRO LEU GLU LYS SEQRES 3BB 194 PRO VAL THR LEU ARG CYS GLN GLY PRO PRO GLY VAL ASP SEQRES 4BB 194 LEU TYR ARG LEU GLU LYS LEU SER SER SER ARG TYR GLN SEQRES 5BB 194 ASP GLN ALA VAL LEU PHE ILE PRO ALA MET LYS ARG SER SEQRES 6BB 194 LEU ALA GLY ARG TYR ARG CYS SER TYR GLN GLN GLY SER SEQRES 7BB 194 LEU TRP SER LEU PRO SER ASP GLN LEU GLU LEU VAL ALA SEQRES 8BB 194 THR GLY VAL PHE ALA LYS PRO SER LEU SER ALA GLN PRO SEQRES 9BB 194 GLY PRO ALA VAL SER SER GLY GLY ASP VAL THR LEU GLN SEQRES 10BB 194 CYS GLN THR ARG TYR GLY PHE ASP GLN PHE ALA LEU TYR SEQRES 11BB 194 LYS GLU GLY ASP PRO ALA PRO TYR LYS ASN PRO GLU ARG SEQRES 12BB 194 TRP TYR ARG ALA SER PHE PRO ILE ILE THR VAL THR ALA SEQRES 13BB 194 ALA HIS SER GLY THR TYR ARG CYS TYR SER PHE SER SER SEQRES 14BB 194 ARG ASP PRO TYR LEU TRP SER ALA PRO SER ASP PRO LEU SEQRES 15BB 194 GLU LEU VAL VAL THR GLY ASP PRO ILE GLU GLY ARG SEQRES 1DD 130 GLN VAL GLN LEU GLN GLU SER GLY GLY GLY LEU VAL GLN SEQRES 2DD 130 PRO GLY GLY SER LEU ARG LEU SER CYS ALA ALA ALA GLY SEQRES 3DD 130 PHE THR PHE ASP TYR TYR ALA ILE ALA TRP PHE ARG GLN SEQRES 4DD 130 ALA PRO GLY LYS GLU ARG GLU GLY VAL SER CYS ILE SER SEQRES 5DD 130 SER SER ASP GLY THR THR TYR TYR ALA ASP SER VAL LYS SEQRES 6DD 130 GLY ARG PHE THR ILE SER LYS ASP ASN ALA LYS ASN THR SEQRES 7DD 130 MET TYR LEU GLN MET ASN SER LEU LYS PRO GLU ASP THR SEQRES 8DD 130 ALA VAL TYR TYR CYS ALA THR SER PRO LEU TYR SER THR SEQRES 9DD 130 ASN ASP ARG CYS ILE SER GLU ASP TYR ASP TYR TRP GLY SEQRES 10DD 130 GLN GLY THR GLN VAL THR VAL SER SER LEU VAL PRO ARG SEQRES 1CC 130 GLN VAL GLN LEU GLN GLU SER GLY GLY GLY LEU VAL GLN SEQRES 2CC 130 PRO GLY GLY SER LEU ARG LEU SER CYS ALA ALA ALA GLY SEQRES 3CC 130 PHE THR PHE ASP TYR TYR ALA ILE ALA TRP PHE ARG GLN SEQRES 4CC 130 ALA PRO GLY LYS GLU ARG GLU GLY VAL SER CYS ILE SER SEQRES 5CC 130 SER SER ASP GLY THR THR TYR TYR ALA ASP SER VAL LYS SEQRES 6CC 130 GLY ARG PHE THR ILE SER LYS ASP ASN ALA LYS ASN THR SEQRES 7CC 130 MET TYR LEU GLN MET ASN SER LEU LYS PRO GLU ASP THR SEQRES 8CC 130 ALA VAL TYR TYR CYS ALA THR SER PRO LEU TYR SER THR SEQRES 9CC 130 ASN ASP ARG CYS ILE SER GLU ASP TYR ASP TYR TRP GLY SEQRES 10CC 130 GLN GLY THR GLN VAL THR VAL SER SER LEU VAL PRO ARG HET CA CC 201 1 HETNAM CA CALCIUM ION FORMUL 5 CA CA 2+ FORMUL 6 HOH *269(H2 O) HELIX 1 AA1 LYSAA 63 ALAAA 67 5 5 HELIX 2 AA2 THRAA 155 SERAA 159 5 5 HELIX 3 AA3 LYSBB 63 ALABB 67 5 5 HELIX 4 AA4 THRBB 155 SERBB 159 5 5 HELIX 5 AA5 LYSDD 87 THRDD 91 5 5 HELIX 6 AA6 ILEDD 109 TYRDD 113 5 5 HELIX 7 AA7 ASPCC 62 LYSCC 65 5 4 HELIX 8 AA8 LYSCC 87 THRCC 91 5 5 HELIX 9 AA9 ILECC 109 TYRCC 113 5 5 SHEET 1 AA1 3 SERAA 13 LEUAA 17 0 SHEET 2 AA1 3 VALAA 28 GLNAA 33 -1 O THRAA 29 N LEUAA 17 SHEET 3 AA1 3 VALAA 56 ILEAA 59 -1 O ILEAA 59 N VALAA 28 SHEET 1 AA2 4 LEUAA 21 PROAA 23 0 SHEET 2 AA2 4 LEUAA 87 THRAA 92 1 O VALAA 90 N VALAA 22 SHEET 3 AA2 4 GLYAA 68 GLNAA 76 -1 N GLYAA 68 O LEUAA 89 SHEET 4 AA2 4 LEUAA 79 TRPAA 80 -1 O LEUAA 79 N GLNAA 76 SHEET 1 AA3 6 LEUAA 21 PROAA 23 0 SHEET 2 AA3 6 LEUAA 87 THRAA 92 1 O VALAA 90 N VALAA 22 SHEET 3 AA3 6 GLYAA 68 GLNAA 76 -1 N GLYAA 68 O LEUAA 89 SHEET 4 AA3 6 LEUAA 40 LYSAA 45 -1 N GLUAA 44 O ARGAA 71 SHEET 5 AA3 6 ARGAA 50 GLNAA 52 -1 O ARGAA 50 N LYSAA 45 SHEET 6 AA3 6 TYRCC 102 SERCC 103 -1 O TYRCC 102 N TYRAA 51 SHEET 1 AA4 3 SERAA 99 GLNAA 103 0 SHEET 2 AA4 3 VALAA 114 GLNAA 119 -1 O GLNAA 117 N SERAA 101 SHEET 3 AA4 3 SERBB 148 ILEBB 151 -1 O PHEBB 149 N LEUAA 116 SHEET 1 AA5 5 ALAAA 107 SERAA 109 0 SHEET 2 AA5 5 LEUBB 182 THRBB 187 1 O VALBB 185 N VALAA 108 SHEET 3 AA5 5 GLYBB 160 PHEBB 167 -1 N GLYBB 160 O LEUBB 184 SHEET 4 AA5 5 GLNAA 126 LYSAA 131 -1 N TYRAA 130 O ARGBB 163 SHEET 5 AA5 5 ALABB 136 PROBB 137 -1 O ALABB 136 N LYSAA 131 SHEET 1 AA6 5 ALAAA 136 PROAA 137 0 SHEET 2 AA6 5 GLNBB 126 LYSBB 131 -1 O LYSBB 131 N ALAAA 136 SHEET 3 AA6 5 GLYAA 160 PHEAA 167 -1 N ARGAA 163 O TYRBB 130 SHEET 4 AA6 5 LEUAA 182 THRAA 187 -1 O LEUAA 184 N GLYAA 160 SHEET 5 AA6 5 ALABB 107 SERBB 109 1 O VALBB 108 N VALAA 185 SHEET 1 AA7 3 SERAA 148 ILEAA 151 0 SHEET 2 AA7 3 VALBB 114 GLNBB 119 -1 O VALBB 114 N ILEAA 151 SHEET 3 AA7 3 SERBB 99 GLNBB 103 -1 N GLNBB 103 O THRBB 115 SHEET 1 AA8 3 SERBB 13 LEUBB 17 0 SHEET 2 AA8 3 VALBB 28 GLNBB 33 -1 O GLNBB 33 N SERBB 13 SHEET 3 AA8 3 VALBB 56 ILEBB 59 -1 O ILEBB 59 N VALBB 28 SHEET 1 AA9 4 LEUBB 21 PROBB 23 0 SHEET 2 AA9 4 LEUBB 87 THRBB 92 1 O VALBB 90 N VALBB 22 SHEET 3 AA9 4 GLYBB 68 GLNBB 76 -1 N GLYBB 68 O LEUBB 89 SHEET 4 AA9 4 LEUBB 79 TRPBB 80 -1 O LEUBB 79 N GLNBB 76 SHEET 1 AB1 6 LEUBB 21 PROBB 23 0 SHEET 2 AB1 6 LEUBB 87 THRBB 92 1 O VALBB 90 N VALBB 22 SHEET 3 AB1 6 GLYBB 68 GLNBB 76 -1 N GLYBB 68 O LEUBB 89 SHEET 4 AB1 6 LEUBB 40 LYSBB 45 -1 N GLUBB 44 O ARGBB 71 SHEET 5 AB1 6 ARGBB 50 GLNBB 52 -1 O GLNBB 52 N LEUBB 43 SHEET 6 AB1 6 TYRDD 102 SERDD 103 -1 O TYRDD 102 N TYRBB 51 SHEET 1 AB2 4 GLNDD 3 SERDD 7 0 SHEET 2 AB2 4 LEUDD 18 ALADD 25 -1 O ALADD 25 N GLNDD 3 SHEET 3 AB2 4 THRDD 78 METDD 83 -1 O METDD 83 N LEUDD 18 SHEET 4 AB2 4 PHEDD 68 ASPDD 73 -1 N THRDD 69 O GLNDD 82 SHEET 1 AB3 6 GLYDD 10 VALDD 12 0 SHEET 2 AB3 6 THRDD 120 VALDD 124 1 O THRDD 123 N GLYDD 10 SHEET 3 AB3 6 ALADD 92 SERDD 99 -1 N ALADD 92 O VALDD 122 SHEET 4 AB3 6 ALADD 33 GLNDD 39 -1 N PHEDD 37 O TYRDD 95 SHEET 5 AB3 6 GLUDD 46 ILEDD 51 -1 O GLUDD 46 N ARGDD 38 SHEET 6 AB3 6 TYRDD 59 TYRDD 60 -1 O TYRDD 59 N CYSDD 50 SHEET 1 AB4 4 GLYDD 10 VALDD 12 0 SHEET 2 AB4 4 THRDD 120 VALDD 124 1 O THRDD 123 N GLYDD 10 SHEET 3 AB4 4 ALADD 92 SERDD 99 -1 N ALADD 92 O VALDD 122 SHEET 4 AB4 4 TYRDD 115 TRPDD 116 -1 O TYRDD 115 N THRDD 98 SHEET 1 AB5 4 GLNCC 3 SERCC 7 0 SHEET 2 AB5 4 LEUCC 18 ALACC 25 -1 O ALACC 25 N GLNCC 3 SHEET 3 AB5 4 THRCC 78 METCC 83 -1 O METCC 83 N LEUCC 18 SHEET 4 AB5 4 PHECC 68 ASPCC 73 -1 N SERCC 71 O TYRCC 80 SHEET 1 AB6 6 GLYCC 10 VALCC 12 0 SHEET 2 AB6 6 THRCC 120 VALCC 124 1 O THRCC 123 N GLYCC 10 SHEET 3 AB6 6 ALACC 92 SERCC 99 -1 N TYRCC 94 O THRCC 120 SHEET 4 AB6 6 ALACC 33 GLNCC 39 -1 N ALACC 35 O ALACC 97 SHEET 5 AB6 6 GLUCC 46 ILECC 51 -1 O VALCC 48 N TRPCC 36 SHEET 6 AB6 6 TYRCC 59 TYRCC 60 -1 O TYRCC 59 N CYSCC 50 SHEET 1 AB7 4 GLYCC 10 VALCC 12 0 SHEET 2 AB7 4 THRCC 120 VALCC 124 1 O THRCC 123 N GLYCC 10 SHEET 3 AB7 4 ALACC 92 SERCC 99 -1 N TYRCC 94 O THRCC 120 SHEET 4 AB7 4 TYRCC 115 TRPCC 116 -1 O TYRCC 115 N THRCC 98 SSBOND 1 CYSAA 32 CYSAA 72 1555 1555 2.05 SSBOND 2 CYSAA 118 CYSBB 164 1555 1555 2.10 SSBOND 3 CYSAA 164 CYSBB 118 1555 1555 2.09 SSBOND 4 CYSBB 32 CYSBB 72 1555 1555 2.06 SSBOND 5 CYSDD 22 CYSDD 96 1555 1555 2.09 SSBOND 6 CYSDD 50 CYSDD 108 1555 1555 2.13 SSBOND 7 CYSCC 22 CYSCC 96 1555 1555 2.07 SSBOND 8 CYSCC 50 CYSCC 108 1555 1555 2.11 LINK OE1 GLUDD 6 CA CACC 201 1555 2454 2.45 LINK O GLYDD 119 CA CACC 201 1555 2454 2.39 LINK O HOHDD 219 CA CACC 201 2455 1555 2.08 LINK OE1 GLUCC 6 CA CACC 201 1555 1555 2.43 LINK O GLYCC 119 CA CACC 201 1555 1555 2.36 LINK CA CACC 201 O HOHCC 310 1555 1555 2.39 CISPEP 1 LEUAA 17 PROAA 18 0 0.00 CISPEP 2 GLNAA 103 PROAA 104 0 5.77 CISPEP 3 LEUBB 17 PROBB 18 0 8.23 CISPEP 4 GLNBB 103 PROBB 104 0 5.83 CRYST1 69.906 84.751 124.041 90.00 90.00 90.00 P 21 21 21 8 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 0.014305 0.000000 0.000000 0.00000 SCALE2 0.000000 0.011799 0.000000 0.00000 SCALE3 0.000000 0.000000 0.008062 0.00000