HEADER IMMUNE SYSTEM 11-OCT-23 8UJI TITLE ANTI-PHOSPHOHISTIDINE FAB HSC44.CK WITH 3PTZA PEPTIDE COMPND MOL_ID: 1; COMPND 2 MOLECULE: HSC44.CK FAB HEAVY CHAIN; COMPND 3 CHAIN: H, A; COMPND 4 ENGINEERED: YES; COMPND 5 MOL_ID: 2; COMPND 6 MOLECULE: HSC44.CK FAB LIGHT CHAIN; COMPND 7 CHAIN: L, B; COMPND 8 ENGINEERED: YES; COMPND 9 MOL_ID: 3; COMPND 10 MOLECULE: 3-PTZA PEPTIDE; COMPND 11 CHAIN: C, D; COMPND 12 ENGINEERED: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: ORYCTOLAGUS CUNICULUS; SOURCE 3 ORGANISM_TAXID: 9986; SOURCE 4 EXPRESSION_SYSTEM: HOMO SAPIENS; SOURCE 5 EXPRESSION_SYSTEM_TAXID: 9606; SOURCE 6 EXPRESSION_SYSTEM_CELL_LINE: EXPI293F; SOURCE 7 MOL_ID: 2; SOURCE 8 ORGANISM_SCIENTIFIC: ORYCTOLAGUS CUNICULUS; SOURCE 9 ORGANISM_TAXID: 9986; SOURCE 10 EXPRESSION_SYSTEM: HOMO SAPIENS; SOURCE 11 EXPRESSION_SYSTEM_TAXID: 9606; SOURCE 12 EXPRESSION_SYSTEM_CELL_LINE: EXPI293F; SOURCE 13 MOL_ID: 3; SOURCE 14 SYNTHETIC: YES; SOURCE 15 ORGANISM_SCIENTIFIC: SYNTHETIC CONSTRUCT; SOURCE 16 ORGANISM_TAXID: 32630 KEYWDS ANTI-PHOSPHOHISTIDINE ANTIBODY POST-TRANSLATIONAL MODIFICATION KEYWDS 2 ANTIBODY HUMANIZATION, IMMUNE SYSTEM EXPDTA X-RAY DIFFRACTION AUTHOR R.KALAGIRI,R.L.STANFIELD,T.HUNTER,I.A.WILSON REVDAT 1 19-MAR-25 8UJI 0 JRNL AUTH G.D.MARTYN,R.KALAGIRI,G.VEGGIANI,R.L.STANFIELD,I.CHOUDHURI, JRNL AUTH 2 M.SALA,J.MEISENHELDER,C.CHEN,A.BISWAS,R.M.LEVY,D.LYUMKIS, JRNL AUTH 3 I.A.WILSON,T.HUNTER,S.S.SIDHU JRNL TITL USING PHAGE DISPLAY FOR RATIONAL ENGINEERING OF A HIGHER JRNL TITL 2 AFFINITY HUMANIZED 3' PHOSPHOHISTIDINE-SPECIFIC ANTIBODY. JRNL REF BIORXIV 2024 JRNL REFN ISSN 2692-8205 JRNL PMID 39574610 JRNL DOI 10.1101/2024.11.04.621849 REMARK 2 REMARK 2 RESOLUTION. 1.95 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : PHENIX 1.19.2_4158 REMARK 3 AUTHORS : PAUL ADAMS,PAVEL AFONINE,VINCENT CHEN,IAN REMARK 3 : DAVIS,KRESHNA GOPAL,RALF GROSSE-KUNSTLEVE, REMARK 3 : LI-WEI HUNG,ROBERT IMMORMINO,TOM IOERGER, REMARK 3 : AIRLIE MCCOY,ERIK MCKEE,NIGEL MORIARTY, REMARK 3 : REETAL PAI,RANDY READ,JANE RICHARDSON, REMARK 3 : DAVID RICHARDSON,TOD ROMO,JIM SACCHETTINI, REMARK 3 : NICHOLAS SAUTER,JACOB SMITH,LAURENT REMARK 3 : STORONI,TOM TERWILLIGER,PETER ZWART REMARK 3 REMARK 3 REFINEMENT TARGET : GEOSTD + MONOMER LIBRARY + CDL V1.2 REMARK 3 REMARK 3 DATA USED IN REFINEMENT. REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 1.95 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 39.34 REMARK 3 MIN(FOBS/SIGMA_FOBS) : 1.340 REMARK 3 COMPLETENESS FOR RANGE (%) : 97.2 REMARK 3 NUMBER OF REFLECTIONS : 68016 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 R VALUE (WORKING + TEST SET) : 0.214 REMARK 3 R VALUE (WORKING SET) : 0.212 REMARK 3 FREE R VALUE : 0.260 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 4.990 REMARK 3 FREE R VALUE TEST SET COUNT : 3393 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT (IN BINS). REMARK 3 BIN RESOLUTION RANGE COMPL. NWORK NFREE RWORK RFREE REMARK 3 1 39.3400 - 5.6300 0.95 2853 128 0.1670 0.1805 REMARK 3 2 5.6300 - 4.4700 0.99 2831 160 0.1550 0.1889 REMARK 3 3 4.4700 - 3.9000 1.00 2801 144 0.1689 0.2038 REMARK 3 4 3.9000 - 3.5500 0.93 2628 137 0.2008 0.2752 REMARK 3 5 3.5500 - 3.2900 0.97 2711 137 0.2079 0.2668 REMARK 3 6 3.2900 - 3.1000 0.98 2701 153 0.2107 0.2702 REMARK 3 7 3.1000 - 2.9400 0.99 2730 167 0.2274 0.2884 REMARK 3 8 2.9400 - 2.8200 0.99 2734 144 0.2304 0.2472 REMARK 3 9 2.8200 - 2.7100 0.99 2738 156 0.2291 0.2692 REMARK 3 10 2.7100 - 2.6100 0.99 2723 148 0.2311 0.2966 REMARK 3 11 2.6100 - 2.5300 0.99 2707 144 0.2348 0.2840 REMARK 3 12 2.5300 - 2.4600 0.92 2535 142 0.2422 0.3010 REMARK 3 13 2.4600 - 2.4000 0.97 2672 153 0.2415 0.3251 REMARK 3 14 2.4000 - 2.3400 0.99 2735 132 0.2429 0.2975 REMARK 3 15 2.3400 - 2.2800 0.99 2703 143 0.2595 0.3327 REMARK 3 16 2.2800 - 2.2300 0.98 2687 129 0.2500 0.2587 REMARK 3 17 2.2300 - 2.1900 0.99 2723 140 0.2528 0.3216 REMARK 3 18 2.1900 - 2.1500 0.99 2713 133 0.2630 0.3668 REMARK 3 19 2.1500 - 2.1100 0.99 2724 141 0.2585 0.3157 REMARK 3 20 2.1100 - 2.0700 0.99 2709 143 0.2571 0.2813 REMARK 3 21 2.0700 - 2.0400 0.98 2672 136 0.2674 0.3192 REMARK 3 22 2.0400 - 2.0100 0.97 2644 148 0.2865 0.3326 REMARK 3 23 2.0100 - 1.9800 0.97 2675 125 0.3200 0.4074 REMARK 3 24 1.9800 - 1.9500 0.83 2274 110 0.3499 0.3975 REMARK 3 REMARK 3 BULK SOLVENT MODELLING. REMARK 3 METHOD USED : FLAT BULK SOLVENT MODEL REMARK 3 SOLVENT RADIUS : 1.11 REMARK 3 SHRINKAGE RADIUS : 0.90 REMARK 3 K_SOL : NULL REMARK 3 B_SOL : NULL REMARK 3 REMARK 3 ERROR ESTIMATES. REMARK 3 COORDINATE ERROR (MAXIMUM-LIKELIHOOD BASED) : 0.285 REMARK 3 PHASE ERROR (DEGREES, MAXIMUM-LIKELIHOOD BASED) : 28.518 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : 29.81 REMARK 3 MEAN B VALUE (OVERALL, A**2) : 37.81 REMARK 3 OVERALL ANISOTROPIC B VALUE. REMARK 3 B11 (A**2) : NULL REMARK 3 B22 (A**2) : NULL REMARK 3 B33 (A**2) : NULL REMARK 3 B12 (A**2) : NULL REMARK 3 B13 (A**2) : NULL REMARK 3 B23 (A**2) : NULL REMARK 3 REMARK 3 TWINNING INFORMATION. REMARK 3 FRACTION: NULL REMARK 3 OPERATOR: NULL REMARK 3 REMARK 3 DEVIATIONS FROM IDEAL VALUES. REMARK 3 RMSD COUNT REMARK 3 BOND : 0.006 6765 REMARK 3 ANGLE : 0.914 9206 REMARK 3 CHIRALITY : 0.053 1040 REMARK 3 PLANARITY : 0.006 1174 REMARK 3 DIHEDRAL : 13.909 2397 REMARK 3 REMARK 3 TLS DETAILS REMARK 3 NUMBER OF TLS GROUPS : NULL REMARK 3 REMARK 3 NCS DETAILS REMARK 3 NUMBER OF NCS GROUPS : NULL REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: NULL REMARK 4 REMARK 4 8UJI COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 31-OCT-23. REMARK 100 THE DEPOSITION ID IS D_1000277248. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 19-JUL-22 REMARK 200 TEMPERATURE (KELVIN) : 100 REMARK 200 PH : NULL REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : Y REMARK 200 RADIATION SOURCE : SSRL REMARK 200 BEAMLINE : BL12-1 REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 0.97946 REMARK 200 MONOCHROMATOR : NULL REMARK 200 OPTICS : NULL REMARK 200 REMARK 200 DETECTOR TYPE : PIXEL REMARK 200 DETECTOR MANUFACTURER : DECTRIS EIGER2 XE 16M REMARK 200 INTENSITY-INTEGRATION SOFTWARE : HKL-2000 REMARK 200 DATA SCALING SOFTWARE : HKL-2000 REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 68051 REMARK 200 RESOLUTION RANGE HIGH (A) : 1.950 REMARK 200 RESOLUTION RANGE LOW (A) : 39.340 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : -3.000 REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 97.5 REMARK 200 DATA REDUNDANCY : 10.80 REMARK 200 R MERGE (I) : 0.14300 REMARK 200 R SYM (I) : 0.14300 REMARK 200 FOR THE DATA SET : 17.6000 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) : 1.98 REMARK 200 COMPLETENESS FOR SHELL (%) : 91.3 REMARK 200 DATA REDUNDANCY IN SHELL : 7.80 REMARK 200 R MERGE FOR SHELL (I) : 1.93000 REMARK 200 R SYM FOR SHELL (I) : 1.93000 REMARK 200 FOR SHELL : 1.200 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: NULL REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 50.18 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.47 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 0.2M LITHIUM CITRATE, 20% PEG-3350, REMARK 280 VAPOR DIFFUSION, SITTING DROP, TEMPERATURE 293K REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 21 21 2 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 46.24900 REMARK 290 SMTRY2 3 0.000000 1.000000 0.000000 69.80750 REMARK 290 SMTRY3 3 0.000000 0.000000 -1.000000 0.00000 REMARK 290 SMTRY1 4 1.000000 0.000000 0.000000 46.24900 REMARK 290 SMTRY2 4 0.000000 -1.000000 0.000000 69.80750 REMARK 290 SMTRY3 4 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: TRIMERIC REMARK 350 APPLY THE FOLLOWING TO CHAINS: H, L, 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 350 REMARK 350 BIOMOLECULE: 2 REMARK 350 AUTHOR DETERMINED BIOLOGICAL UNIT: TRIMERIC REMARK 350 APPLY THE FOLLOWING TO CHAINS: A, B, 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 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 GLU H 1 REMARK 465 SER H 215 REMARK 465 CYS H 216 REMARK 465 ASP H 217 REMARK 465 LYS H 218 REMARK 465 THR H 219 REMARK 465 HIS H 220 REMARK 465 THR H 221 REMARK 465 GLU A 1 REMARK 465 SER A 215 REMARK 465 CYS A 216 REMARK 465 ASP A 217 REMARK 465 LYS A 218 REMARK 465 THR A 219 REMARK 465 HIS A 220 REMARK 465 THR A 221 REMARK 465 ALA C 1 REMARK 465 GLY C 2 REMARK 465 ALA C 3 REMARK 465 GLY C 9 REMARK 465 ALA D 1 REMARK 465 GLY D 2 REMARK 465 ALA D 8 REMARK 465 GLY D 9 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 CYS H 140 CA - CB - SG ANGL. DEV. = 8.8 DEGREES REMARK 500 CYS A 140 CA - CB - SG ANGL. DEV. = 9.0 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 ILE H 48 -61.60 -91.96 REMARK 500 ARG L 29 16.32 58.70 REMARK 500 ILE L 51 -43.11 70.55 REMARK 500 SER L 94 -164.24 -122.21 REMARK 500 LEU A 4 94.86 -162.18 REMARK 500 SER A 130 -3.20 62.51 REMARK 500 THR A 131 -101.34 39.02 REMARK 500 SER A 132 105.22 3.12 REMARK 500 ARG B 29 19.10 56.61 REMARK 500 LYS B 31 -26.98 -146.04 REMARK 500 ILE B 51 -48.84 73.10 REMARK 500 PHE B 83 101.81 -54.25 REMARK 500 SER B 94 -167.15 -117.12 REMARK 500 REMARK 500 REMARK: NULL DBREF 8UJI H 1 221 PDB 8UJI 8UJI 1 221 DBREF 8UJI L 1 212 PDB 8UJI 8UJI 1 212 DBREF 8UJI A 1 221 PDB 8UJI 8UJI 1 221 DBREF 8UJI B 1 212 PDB 8UJI 8UJI 1 212 DBREF 8UJI C 1 9 PDB 8UJI 8UJI 1 9 DBREF 8UJI D 1 9 PDB 8UJI 8UJI 1 9 SEQRES 1 H 225 GLU VAL GLN LEU VAL GLU SER GLY GLY GLY LEU VAL GLN SEQRES 2 H 225 PRO GLY GLY SER LEU ARG LEU SER CYS THR ALA SER GLY SEQRES 3 H 225 PHE SER ILE ASP SER TYR GLY PHE SER TRP VAL ARG GLN SEQRES 4 H 225 ALA PRO GLY LYS GLY LEU GLU HIS ILE GLY TYR LEU THR SEQRES 5 H 225 ALA GLY GLY ARG ALA PHE TYR ALA SER TRP ALA LYS SER SEQRES 6 H 225 ARG SER THR ILE THR ARG ASN THR ASN GLU ASN THR VAL SEQRES 7 H 225 THR LEU LYS MET ASN SER LEU ARG ALA GLU ASP THR ALA SEQRES 8 H 225 VAL TYR TYR CYS ALA LYS LEU GLY SER GLY ASN PRO VAL SEQRES 9 H 225 ALA ILE TRP GLY GLN GLY THR LEU VAL THR VAL SER SER SEQRES 10 H 225 ALA SER THR LYS GLY PRO SER VAL PHE PRO LEU ALA PRO SEQRES 11 H 225 SER SER LYS SER THR SER GLY GLY THR ALA ALA LEU GLY SEQRES 12 H 225 CYS LEU VAL LYS ASP TYR PHE PRO GLU PRO VAL THR VAL SEQRES 13 H 225 SER TRP ASN SER GLY ALA LEU THR SER GLY VAL HIS THR SEQRES 14 H 225 PHE PRO ALA VAL LEU GLN SER SER GLY LEU TYR SER LEU SEQRES 15 H 225 SER SER VAL VAL THR VAL PRO SER SER SER LEU GLY THR SEQRES 16 H 225 GLN THR TYR ILE CYS ASN VAL ASN HIS LYS PRO SER ASN SEQRES 17 H 225 THR LYS VAL ASP LYS LYS VAL GLU PRO LYS SER CYS ASP SEQRES 18 H 225 LYS THR HIS THR SEQRES 1 L 218 ASP ILE GLN MET THR GLN SER PRO SER SER LEU SER ALA SEQRES 2 L 218 SER VAL GLY ASP ARG VAL THR ILE THR CYS ARG ALA SER SEQRES 3 L 218 GLN SER VAL TRP ARG ASN LYS ASN VAL ALA TRP TYR GLN SEQRES 4 L 218 GLN LYS PRO GLY LYS ALA PRO LYS ARG LEU ILE TYR ALA SEQRES 5 L 218 ILE ALA SER LEU TYR SER GLY VAL PRO SER ARG PHE SER SEQRES 6 L 218 GLY SER GLY SER GLY THR ASP PHE THR LEU THR ILE SER SEQRES 7 L 218 SER LEU GLN PRO GLU ASP PHE ALA THR TYR TYR CYS VAL SEQRES 8 L 218 GLY HIS TYR GLY SER GLU ASN ASP ALA TYR TYR ALA PHE SEQRES 9 L 218 GLY GLN GLY THR LYS VAL GLU ILE LYS ARG THR VAL ALA SEQRES 10 L 218 ALA PRO SER VAL PHE ILE PHE PRO PRO SER ASP GLU GLN SEQRES 11 L 218 LEU LYS SER GLY THR ALA SER VAL VAL CYS LEU LEU ASN SEQRES 12 L 218 ASN PHE TYR PRO ARG GLU ALA LYS VAL SER TRP TYR VAL SEQRES 13 L 218 ASP ASN ALA LEU GLN SER GLY ASN SER GLN GLU SER VAL SEQRES 14 L 218 THR GLU GLN ASP SER LYS ASP SER THR TYR SER LEU SER SEQRES 15 L 218 SER THR LEU THR LEU SER LYS ALA ASP TYR GLU LYS HIS SEQRES 16 L 218 LYS VAL TYR ALA CYS GLU VAL THR GLN GLY THR THR SER SEQRES 17 L 218 VAL THR LYS SER PHE ASN ARG GLY GLU CYS SEQRES 1 A 225 GLU VAL GLN LEU VAL GLU SER GLY GLY GLY LEU VAL GLN SEQRES 2 A 225 PRO GLY GLY SER LEU ARG LEU SER CYS THR ALA SER GLY SEQRES 3 A 225 PHE SER ILE ASP SER TYR GLY PHE SER TRP VAL ARG GLN SEQRES 4 A 225 ALA PRO GLY LYS GLY LEU GLU HIS ILE GLY TYR LEU THR SEQRES 5 A 225 ALA GLY GLY ARG ALA PHE TYR ALA SER TRP ALA LYS SER SEQRES 6 A 225 ARG SER THR ILE THR ARG ASN THR ASN GLU ASN THR VAL SEQRES 7 A 225 THR LEU LYS MET ASN SER LEU ARG ALA GLU ASP THR ALA SEQRES 8 A 225 VAL TYR TYR CYS ALA LYS LEU GLY SER GLY ASN PRO VAL SEQRES 9 A 225 ALA ILE TRP GLY GLN GLY THR LEU VAL THR VAL SER SER SEQRES 10 A 225 ALA SER THR LYS GLY PRO SER VAL PHE PRO LEU ALA PRO SEQRES 11 A 225 SER SER LYS SER THR SER GLY GLY THR ALA ALA LEU GLY SEQRES 12 A 225 CYS LEU VAL LYS ASP TYR PHE PRO GLU PRO VAL THR VAL SEQRES 13 A 225 SER TRP ASN SER GLY ALA LEU THR SER GLY VAL HIS THR SEQRES 14 A 225 PHE PRO ALA VAL LEU GLN SER SER GLY LEU TYR SER LEU SEQRES 15 A 225 SER SER VAL VAL THR VAL PRO SER SER SER LEU GLY THR SEQRES 16 A 225 GLN THR TYR ILE CYS ASN VAL ASN HIS LYS PRO SER ASN SEQRES 17 A 225 THR LYS VAL ASP LYS LYS VAL GLU PRO LYS SER CYS ASP SEQRES 18 A 225 LYS THR HIS THR SEQRES 1 B 218 ASP ILE GLN MET THR GLN SER PRO SER SER LEU SER ALA SEQRES 2 B 218 SER VAL GLY ASP ARG VAL THR ILE THR CYS ARG ALA SER SEQRES 3 B 218 GLN SER VAL TRP ARG ASN LYS ASN VAL ALA TRP TYR GLN SEQRES 4 B 218 GLN LYS PRO GLY LYS ALA PRO LYS ARG LEU ILE TYR ALA SEQRES 5 B 218 ILE ALA SER LEU TYR SER GLY VAL PRO SER ARG PHE SER SEQRES 6 B 218 GLY SER GLY SER GLY THR ASP PHE THR LEU THR ILE SER SEQRES 7 B 218 SER LEU GLN PRO GLU ASP PHE ALA THR TYR TYR CYS VAL SEQRES 8 B 218 GLY HIS TYR GLY SER GLU ASN ASP ALA TYR TYR ALA PHE SEQRES 9 B 218 GLY GLN GLY THR LYS VAL GLU ILE LYS ARG THR VAL ALA SEQRES 10 B 218 ALA PRO SER VAL PHE ILE PHE PRO PRO SER ASP GLU GLN SEQRES 11 B 218 LEU LYS SER GLY THR ALA SER VAL VAL CYS LEU LEU ASN SEQRES 12 B 218 ASN PHE TYR PRO ARG GLU ALA LYS VAL SER TRP TYR VAL SEQRES 13 B 218 ASP ASN ALA LEU GLN SER GLY ASN SER GLN GLU SER VAL SEQRES 14 B 218 THR GLU GLN ASP SER LYS ASP SER THR TYR SER LEU SER SEQRES 15 B 218 SER THR LEU THR LEU SER LYS ALA ASP TYR GLU LYS HIS SEQRES 16 B 218 LYS VAL TYR ALA CYS GLU VAL THR GLN GLY THR THR SER SEQRES 17 B 218 VAL THR LYS SER PHE ASN ARG GLY GLU CYS SEQRES 1 C 9 ALA GLY ALA GLY UKD ALA GLY ALA GLY SEQRES 1 D 9 ALA GLY ALA GLY UKD ALA GLY ALA GLY HET UKD C 5 14 HET UKD D 5 14 HET EDO H 301 4 HETNAM UKD 3-(4-PHOSPHONO-1H-1,2,3-TRIAZOL-1-YL)-L-ALANINE HETNAM EDO 1,2-ETHANEDIOL HETSYN UKD PHIS HETSYN EDO ETHYLENE GLYCOL FORMUL 5 UKD 2(C5 H9 N4 O5 P) FORMUL 7 EDO C2 H6 O2 FORMUL 8 HOH *219(H2 O) HELIX 1 AA1 SER H 28 TYR H 32 5 5 HELIX 2 AA2 SER H 61 LYS H 64 5 4 HELIX 3 AA3 ARG H 83 THR H 87 5 5 HELIX 4 AA4 SER H 156 ALA H 158 5 3 HELIX 5 AA5 SER H 187 LEU H 189 5 3 HELIX 6 AA6 LYS H 201 ASN H 204 5 4 HELIX 7 AA7 VAL L 27B LYS L 31 5 5 HELIX 8 AA8 GLN L 79 PHE L 83 5 5 HELIX 9 AA9 SER L 121 SER L 127 1 7 HELIX 10 AB1 LYS L 183 LYS L 188 1 6 HELIX 11 AB2 SER A 28 TYR A 32 5 5 HELIX 12 AB3 ALA A 60 SER A 65 1 6 HELIX 13 AB4 ARG A 83 THR A 87 5 5 HELIX 14 AB5 SER A 156 ALA A 158 5 3 HELIX 15 AB6 SER A 187 LEU A 189 5 3 HELIX 16 AB7 LYS A 201 ASN A 204 5 4 HELIX 17 AB8 VAL B 27B LYS B 31 5 5 HELIX 18 AB9 GLN B 79 PHE B 83 5 5 HELIX 19 AC1 SER B 121 LYS B 126 1 6 HELIX 20 AC2 LYS B 183 GLU B 187 1 5 SHEET 1 AA1 4 GLN H 3 SER H 7 0 SHEET 2 AA1 4 LEU H 18 SER H 25 -1 O SER H 21 N SER H 7 SHEET 3 AA1 4 THR H 77 MET H 82 -1 O LEU H 80 N LEU H 20 SHEET 4 AA1 4 SER H 67 ASN H 72 -1 N THR H 70 O THR H 79 SHEET 1 AA2 6 GLY H 10 VAL H 12 0 SHEET 2 AA2 6 THR H 107 VAL H 111 1 O THR H 110 N VAL H 12 SHEET 3 AA2 6 ALA H 88 LYS H 94 -1 N ALA H 88 O VAL H 109 SHEET 4 AA2 6 PHE H 34 GLN H 39 -1 N VAL H 37 O TYR H 91 SHEET 5 AA2 6 GLU H 46 LEU H 51 -1 O ILE H 48 N TRP H 36 SHEET 6 AA2 6 ALA H 57 TYR H 59 -1 O PHE H 58 N TYR H 50 SHEET 1 AA3 4 GLY H 10 VAL H 12 0 SHEET 2 AA3 4 THR H 107 VAL H 111 1 O THR H 110 N VAL H 12 SHEET 3 AA3 4 ALA H 88 LYS H 94 -1 N ALA H 88 O VAL H 109 SHEET 4 AA3 4 ILE H 102 TRP H 103 -1 O ILE H 102 N LYS H 94 SHEET 1 AA4 4 SER H 120 LEU H 124 0 SHEET 2 AA4 4 THR H 135 TYR H 145 -1 O LEU H 141 N PHE H 122 SHEET 3 AA4 4 TYR H 176 PRO H 185 -1 O LEU H 178 N VAL H 142 SHEET 4 AA4 4 VAL H 163 THR H 165 -1 N HIS H 164 O VAL H 181 SHEET 1 AA5 4 SER H 120 LEU H 124 0 SHEET 2 AA5 4 THR H 135 TYR H 145 -1 O LEU H 141 N PHE H 122 SHEET 3 AA5 4 TYR H 176 PRO H 185 -1 O LEU H 178 N VAL H 142 SHEET 4 AA5 4 VAL H 169 LEU H 170 -1 N VAL H 169 O SER H 177 SHEET 1 AA6 3 THR H 151 TRP H 154 0 SHEET 2 AA6 3 ILE H 195 HIS H 200 -1 O ASN H 197 N SER H 153 SHEET 3 AA6 3 THR H 205 LYS H 210 -1 O VAL H 207 N VAL H 198 SHEET 1 AA7 4 MET L 4 SER L 7 0 SHEET 2 AA7 4 VAL L 19 ALA L 25 -1 O ARG L 24 N THR L 5 SHEET 3 AA7 4 ASP L 70 ILE L 75 -1 O PHE L 71 N CYS L 23 SHEET 4 AA7 4 PHE L 62 SER L 67 -1 N SER L 65 O THR L 72 SHEET 1 AA8 6 SER L 10 SER L 14 0 SHEET 2 AA8 6 THR L 102 LYS L 107 1 O GLU L 105 N LEU L 11 SHEET 3 AA8 6 ALA L 84 TYR L 92 -1 N ALA L 84 O VAL L 104 SHEET 4 AA8 6 VAL L 33 GLN L 38 -1 N TYR L 36 O TYR L 87 SHEET 5 AA8 6 LYS L 45 TYR L 49 -1 O LEU L 47 N TRP L 35 SHEET 6 AA8 6 SER L 53 LEU L 54 -1 O SER L 53 N TYR L 49 SHEET 1 AA9 4 SER L 10 SER L 14 0 SHEET 2 AA9 4 THR L 102 LYS L 107 1 O GLU L 105 N LEU L 11 SHEET 3 AA9 4 ALA L 84 TYR L 92 -1 N ALA L 84 O VAL L 104 SHEET 4 AA9 4 TYR L 95D PHE L 98 -1 O TYR L 95D N TYR L 92 SHEET 1 AB1 4 SER L 114 PHE L 118 0 SHEET 2 AB1 4 THR L 129 PHE L 139 -1 O LEU L 135 N PHE L 116 SHEET 3 AB1 4 TYR L 173 SER L 182 -1 O LEU L 179 N VAL L 132 SHEET 4 AB1 4 SER L 159 VAL L 163 -1 N SER L 162 O SER L 176 SHEET 1 AB2 4 ALA L 153 LEU L 154 0 SHEET 2 AB2 4 LYS L 145 VAL L 150 -1 N VAL L 150 O ALA L 153 SHEET 3 AB2 4 VAL L 191 GLN L 198 -1 O GLU L 195 N SER L 147 SHEET 4 AB2 4 THR L 201 ASN L 208 -1 O VAL L 203 N VAL L 196 SHEET 1 AB3 4 GLN A 3 GLY A 8 0 SHEET 2 AB3 4 LEU A 18 SER A 25 -1 O SER A 25 N GLN A 3 SHEET 3 AB3 4 THR A 77 MET A 82 -1 O LEU A 80 N LEU A 20 SHEET 4 AB3 4 SER A 67 ASN A 72 -1 N THR A 70 O THR A 79 SHEET 1 AB4 6 GLY A 10 VAL A 12 0 SHEET 2 AB4 6 THR A 107 VAL A 111 1 O THR A 110 N VAL A 12 SHEET 3 AB4 6 ALA A 88 LYS A 94 -1 N TYR A 90 O THR A 107 SHEET 4 AB4 6 PHE A 34 GLN A 39 -1 N SER A 35 O ALA A 93 SHEET 5 AB4 6 GLU A 46 LEU A 51 -1 O LEU A 51 N PHE A 34 SHEET 6 AB4 6 ALA A 57 TYR A 59 -1 O PHE A 58 N TYR A 50 SHEET 1 AB5 4 GLY A 10 VAL A 12 0 SHEET 2 AB5 4 THR A 107 VAL A 111 1 O THR A 110 N VAL A 12 SHEET 3 AB5 4 ALA A 88 LYS A 94 -1 N TYR A 90 O THR A 107 SHEET 4 AB5 4 ILE A 102 TRP A 103 -1 O ILE A 102 N LYS A 94 SHEET 1 AB6 4 SER A 120 LEU A 124 0 SHEET 2 AB6 4 THR A 135 TYR A 145 -1 O GLY A 139 N LEU A 124 SHEET 3 AB6 4 TYR A 176 PRO A 185 -1 O LEU A 178 N VAL A 142 SHEET 4 AB6 4 VAL A 163 THR A 165 -1 N HIS A 164 O VAL A 181 SHEET 1 AB7 4 SER A 120 LEU A 124 0 SHEET 2 AB7 4 THR A 135 TYR A 145 -1 O GLY A 139 N LEU A 124 SHEET 3 AB7 4 TYR A 176 PRO A 185 -1 O LEU A 178 N VAL A 142 SHEET 4 AB7 4 VAL A 169 LEU A 170 -1 N VAL A 169 O SER A 177 SHEET 1 AB8 3 THR A 151 TRP A 154 0 SHEET 2 AB8 3 ILE A 195 HIS A 200 -1 O ASN A 197 N SER A 153 SHEET 3 AB8 3 THR A 205 LYS A 210 -1 O VAL A 207 N VAL A 198 SHEET 1 AB9 4 MET B 4 SER B 7 0 SHEET 2 AB9 4 VAL B 19 ALA B 25 -1 O ARG B 24 N THR B 5 SHEET 3 AB9 4 ASP B 70 ILE B 75 -1 O LEU B 73 N ILE B 21 SHEET 4 AB9 4 PHE B 62 SER B 67 -1 N SER B 63 O THR B 74 SHEET 1 AC1 6 SER B 10 ALA B 13 0 SHEET 2 AC1 6 THR B 102 ILE B 106 1 O GLU B 105 N LEU B 11 SHEET 3 AC1 6 ALA B 84 TYR B 92 -1 N ALA B 84 O VAL B 104 SHEET 4 AC1 6 VAL B 33 GLN B 38 -1 N TYR B 36 O TYR B 87 SHEET 5 AC1 6 LYS B 45 TYR B 49 -1 O LYS B 45 N GLN B 37 SHEET 6 AC1 6 SER B 53 LEU B 54 -1 O SER B 53 N TYR B 49 SHEET 1 AC2 4 SER B 10 ALA B 13 0 SHEET 2 AC2 4 THR B 102 ILE B 106 1 O GLU B 105 N LEU B 11 SHEET 3 AC2 4 ALA B 84 TYR B 92 -1 N ALA B 84 O VAL B 104 SHEET 4 AC2 4 TYR B 95D PHE B 98 -1 O TYR B 95D N TYR B 92 SHEET 1 AC3 4 SER B 114 PHE B 118 0 SHEET 2 AC3 4 THR B 129 PHE B 139 -1 O VAL B 133 N PHE B 118 SHEET 3 AC3 4 TYR B 173 SER B 182 -1 O LEU B 179 N VAL B 132 SHEET 4 AC3 4 SER B 159 VAL B 163 -1 N SER B 162 O SER B 176 SHEET 1 AC4 4 ALA B 153 LEU B 154 0 SHEET 2 AC4 4 LYS B 145 VAL B 150 -1 N VAL B 150 O ALA B 153 SHEET 3 AC4 4 VAL B 191 GLN B 198 -1 O GLU B 195 N SER B 147 SHEET 4 AC4 4 THR B 201 ASN B 208 -1 O VAL B 203 N VAL B 196 SSBOND 1 CYS H 22 CYS H 92 1555 1555 2.02 SSBOND 2 CYS H 140 CYS H 196 1555 1555 2.04 SSBOND 3 CYS L 23 CYS L 88 1555 1555 2.05 SSBOND 4 CYS L 134 CYS L 194 1555 1555 2.02 SSBOND 5 CYS A 22 CYS A 92 1555 1555 2.02 SSBOND 6 CYS A 140 CYS A 196 1555 1555 2.05 SSBOND 7 CYS B 23 CYS B 88 1555 1555 2.06 SSBOND 8 CYS B 134 CYS B 194 1555 1555 2.03 LINK C GLY C 4 N UKD C 5 1555 1555 1.33 LINK C UKD C 5 N ALA C 6 1555 1555 1.33 LINK C GLY D 4 N UKD D 5 1555 1555 1.33 LINK C UKD D 5 N ALA D 6 1555 1555 1.33 CISPEP 1 PHE H 146 PRO H 147 0 -7.68 CISPEP 2 GLU H 148 PRO H 149 0 -0.88 CISPEP 3 SER L 7 PRO L 8 0 -3.58 CISPEP 4 TYR L 140 PRO L 141 0 1.19 CISPEP 5 PHE A 146 PRO A 147 0 -5.89 CISPEP 6 GLU A 148 PRO A 149 0 -4.02 CISPEP 7 SER B 7 PRO B 8 0 -7.90 CISPEP 8 TYR B 140 PRO B 141 0 4.37 CRYST1 92.498 139.615 73.616 90.00 90.00 90.00 P 21 21 2 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 0.010811 0.000000 0.000000 0.00000 SCALE2 0.000000 0.007163 0.000000 0.00000 SCALE3 0.000000 0.000000 0.013584 0.00000