HEADER IMMUNE SYSTEM 09-OCT-23 8UHP TITLE ANTI-PHOSPHOHISTIDINE FAB HSC44.CK.20.N32F WITH 3PTZA PEPTIDE COMPND MOL_ID: 1; COMPND 2 MOLECULE: HSC44.CK.20.N32F FAB HEAVY CHAIN; COMPND 3 CHAIN: H, B; COMPND 4 ENGINEERED: YES; COMPND 5 MOL_ID: 2; COMPND 6 MOLECULE: HSC44.CK.20.N32F FAB LIGHT CHAIN; COMPND 7 CHAIN: L, A; COMPND 8 ENGINEERED: YES; COMPND 9 MOL_ID: 3; COMPND 10 MOLECULE: 3PTZA PEPTIDE; COMPND 11 CHAIN: D, C; 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 MOL_ID: 2; SOURCE 7 ORGANISM_SCIENTIFIC: ORYCTOLAGUS CUNICULUS; SOURCE 8 ORGANISM_TAXID: 9986; SOURCE 9 EXPRESSION_SYSTEM: HOMO SAPIENS; SOURCE 10 EXPRESSION_SYSTEM_TAXID: 9606; SOURCE 11 MOL_ID: 3; SOURCE 12 SYNTHETIC: YES; SOURCE 13 ORGANISM_SCIENTIFIC: SYNTHETIC CONSTRUCT; SOURCE 14 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 8UHP 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. 1.98 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : PHENIX 1.21RC1_5127 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) : 1.98 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 47.25 REMARK 3 MIN(FOBS/SIGMA_FOBS) : 1.340 REMARK 3 COMPLETENESS FOR RANGE (%) : 99.0 REMARK 3 NUMBER OF REFLECTIONS : 88933 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 R VALUE (WORKING + TEST SET) : 0.218 REMARK 3 R VALUE (WORKING SET) : 0.216 REMARK 3 FREE R VALUE : 0.247 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 4.790 REMARK 3 FREE R VALUE TEST SET COUNT : 4259 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 47.2500 - 6.1400 1.00 3108 143 0.1840 0.2285 REMARK 3 2 6.1400 - 4.8700 1.00 2970 142 0.1578 0.1667 REMARK 3 3 4.8700 - 4.2600 1.00 2927 136 0.1476 0.1707 REMARK 3 4 4.2600 - 3.8700 1.00 2891 158 0.1713 0.1761 REMARK 3 5 3.8700 - 3.5900 1.00 2865 143 0.1972 0.2377 REMARK 3 6 3.5900 - 3.3800 1.00 2881 130 0.2115 0.2078 REMARK 3 7 3.3800 - 3.2100 1.00 2837 147 0.2184 0.2489 REMARK 3 8 3.2100 - 3.0700 1.00 2842 170 0.2322 0.2772 REMARK 3 9 3.0700 - 2.9500 1.00 2850 130 0.2312 0.2670 REMARK 3 10 2.9500 - 2.8500 1.00 2816 166 0.2381 0.2738 REMARK 3 11 2.8500 - 2.7600 1.00 2835 150 0.2367 0.2719 REMARK 3 12 2.7600 - 2.6800 1.00 2854 140 0.2379 0.2825 REMARK 3 13 2.6800 - 2.6100 1.00 2841 133 0.2398 0.2696 REMARK 3 14 2.6100 - 2.5500 0.99 2768 148 0.2426 0.2859 REMARK 3 15 2.5500 - 2.4900 0.99 2839 142 0.2384 0.2735 REMARK 3 16 2.4900 - 2.4400 0.99 2831 127 0.2514 0.3189 REMARK 3 17 2.4400 - 2.3900 0.99 2724 181 0.2681 0.3149 REMARK 3 18 2.3900 - 2.3400 0.99 2837 137 0.2557 0.3541 REMARK 3 19 2.3400 - 2.3000 0.99 2793 146 0.2666 0.2836 REMARK 3 20 2.3000 - 2.2600 0.99 2804 131 0.2737 0.3102 REMARK 3 21 2.2600 - 2.2300 0.99 2749 159 0.2730 0.3132 REMARK 3 22 2.2300 - 2.1900 0.99 2786 149 0.2724 0.3238 REMARK 3 23 2.1900 - 2.1600 0.99 2747 146 0.2789 0.3396 REMARK 3 24 2.1600 - 2.1300 0.99 2805 147 0.2855 0.2973 REMARK 3 25 2.1300 - 2.1000 0.98 2724 135 0.2927 0.3346 REMARK 3 26 2.1000 - 2.0700 0.98 2847 129 0.2915 0.3045 REMARK 3 27 2.0700 - 2.0500 0.98 2740 108 0.3021 0.3461 REMARK 3 28 2.0500 - 2.0200 0.98 2785 134 0.2985 0.3195 REMARK 3 29 2.0200 - 2.0000 0.98 2753 124 0.3106 0.3543 REMARK 3 30 2.0000 - 1.9800 0.93 2625 128 0.3145 0.3165 REMARK 3 REMARK 3 BULK SOLVENT MODELLING. REMARK 3 METHOD USED : FLAT BULK SOLVENT MODEL REMARK 3 SOLVENT RADIUS : 1.10 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) : 0.276 REMARK 3 PHASE ERROR (DEGREES, MAXIMUM-LIKELIHOOD BASED) : 26.438 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : 31.95 REMARK 3 MEAN B VALUE (OVERALL, A**2) : 41.99 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 : 0.008 6821 REMARK 3 ANGLE : 0.855 9274 REMARK 3 CHIRALITY : 0.054 1045 REMARK 3 PLANARITY : 0.006 1180 REMARK 3 DIHEDRAL : 13.393 2412 REMARK 3 REMARK 3 TLS DETAILS REMARK 3 NUMBER OF TLS GROUPS : 1 REMARK 3 TLS GROUP : 1 REMARK 3 SELECTION: ALL REMARK 3 ORIGIN FOR THE GROUP (A): 16.9652 -27.8760 -27.8104 REMARK 3 T TENSOR REMARK 3 T11: 0.2027 T22: 0.2670 REMARK 3 T33: 0.2445 T12: -0.0247 REMARK 3 T13: -0.0269 T23: 0.0468 REMARK 3 L TENSOR REMARK 3 L11: 0.0277 L22: 0.5831 REMARK 3 L33: 0.2914 L12: -0.1327 REMARK 3 L13: 0.1053 L23: -0.2967 REMARK 3 S TENSOR REMARK 3 S11: 0.0287 S12: -0.0472 S13: -0.0432 REMARK 3 S21: -0.1126 S22: -0.0511 S23: -0.0370 REMARK 3 S31: 0.0301 S32: 0.0706 S33: 0.0030 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 8UHP COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 04-DEC-23. REMARK 100 THE DEPOSITION ID IS D_1000277439. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 24-SEP-22 REMARK 200 TEMPERATURE (KELVIN) : 100 REMARK 200 PH : 6.5 REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : Y REMARK 200 RADIATION SOURCE : ALS REMARK 200 BEAMLINE : 5.0.1 REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 0.97741 REMARK 200 MONOCHROMATOR : NULL REMARK 200 OPTICS : NULL REMARK 200 REMARK 200 DETECTOR TYPE : PIXEL REMARK 200 DETECTOR MANUFACTURER : DECTRIS PILATUS3 2M REMARK 200 INTENSITY-INTEGRATION SOFTWARE : HKL-2000 REMARK 200 DATA SCALING SOFTWARE : HKL-2000 REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 89758 REMARK 200 RESOLUTION RANGE HIGH (A) : 1.977 REMARK 200 RESOLUTION RANGE LOW (A) : 47.298 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : -3.000 REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 100.0 REMARK 200 DATA REDUNDANCY : 6.000 REMARK 200 R MERGE (I) : 0.12500 REMARK 200 R SYM (I) : 0.12500 REMARK 200 FOR THE DATA SET : 12.9000 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 1.98 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 2.01 REMARK 200 COMPLETENESS FOR SHELL (%) : 100.0 REMARK 200 DATA REDUNDANCY IN SHELL : 5.60 REMARK 200 R MERGE FOR SHELL (I) : 1.35000 REMARK 200 R SYM FOR SHELL (I) : 1.35000 REMARK 200 FOR SHELL : 1.250 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 (%): 63.40 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 3.36 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 0.08M SODIUM CACODYLATE, PH 6.5, 0.16M REMARK 280 CALCIUM ACETATE, 20% GLYCEROL, 14.4% PEG8000, VAPOR DIFFUSION, REMARK 280 SITTING DROP, TEMPERATURE 293K 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 35.77300 REMARK 290 SMTRY2 2 0.000000 -1.000000 0.000000 0.00000 REMARK 290 SMTRY3 2 0.000000 0.000000 1.000000 119.82400 REMARK 290 SMTRY1 3 -1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 3 0.000000 1.000000 0.000000 36.97650 REMARK 290 SMTRY3 3 0.000000 0.000000 -1.000000 119.82400 REMARK 290 SMTRY1 4 1.000000 0.000000 0.000000 35.77300 REMARK 290 SMTRY2 4 0.000000 -1.000000 0.000000 36.97650 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, 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: TRIMERIC REMARK 350 APPLY THE FOLLOWING TO CHAINS: B, A, 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 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 B 1 REMARK 465 SER B 215 REMARK 465 CYS B 216 REMARK 465 ASP B 217 REMARK 465 LYS B 218 REMARK 465 THR B 219 REMARK 465 HIS B 220 REMARK 465 THR B 221 REMARK 465 ASP A 1 REMARK 465 CYS A 212 REMARK 465 ALA D 1 REMARK 465 GLY D 2 REMARK 465 ALA D 3 REMARK 465 GLY D 9 REMARK 465 ALA C 1 REMARK 465 GLY C 2 REMARK 465 ALA C 3 REMARK 465 GLY C 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. = 8.1 DEGREES REMARK 500 CYS L 194 CA - CB - SG ANGL. DEV. = 7.7 DEGREES REMARK 500 CYS B 140 CA - CB - SG ANGL. DEV. = 10.2 DEGREES REMARK 500 CYS A 194 CA - CB - SG ANGL. DEV. = 7.0 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 ASN H 76 30.19 70.38 REMARK 500 SER H 132 50.04 -143.64 REMARK 500 SER H 215 -166.17 -77.41 REMARK 500 ASN L 30 19.24 53.52 REMARK 500 ILE L 51 -55.94 74.25 REMARK 500 ALA L 84 -179.53 -179.47 REMARK 500 ASN B 76 60.07 63.75 REMARK 500 PHE B 146 140.70 -170.89 REMARK 500 ASN A 30 27.16 49.88 REMARK 500 LYS A 31 -23.47 -142.44 REMARK 500 ILE A 51 -51.32 75.23 REMARK 500 SER A 67 108.17 -161.23 REMARK 500 ALA A 84 171.99 177.91 REMARK 500 SER A 94 -166.74 -126.76 REMARK 500 SER A 168 0.07 -67.97 REMARK 500 REMARK 500 REMARK: NULL DBREF 8UHP H 1 221 PDB 8UHP 8UHP 1 221 DBREF 8UHP L 1 212 PDB 8UHP 8UHP 1 212 DBREF 8UHP B 1 221 PDB 8UHP 8UHP 1 221 DBREF 8UHP A 1 212 PDB 8UHP 8UHP 1 212 DBREF 8UHP D 1 9 PDB 8UHP 8UHP 1 9 DBREF 8UHP C 1 9 PDB 8UHP 8UHP 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 THR GLY SER ARG PHE 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 PHE 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 B 225 GLU VAL GLN LEU VAL GLU SER GLY GLY GLY LEU VAL GLN SEQRES 2 B 225 PRO GLY GLY SER LEU ARG LEU SER CYS THR ALA SER GLY SEQRES 3 B 225 PHE SER ILE ASP SER TYR GLY PHE SER TRP VAL ARG GLN SEQRES 4 B 225 ALA PRO GLY LYS GLY LEU GLU HIS ILE GLY TYR LEU THR SEQRES 5 B 225 ALA GLY GLY ARG ALA PHE TYR ALA SER TRP ALA LYS SER SEQRES 6 B 225 ARG SER THR ILE THR ARG ASN THR ASN GLU ASN THR VAL SEQRES 7 B 225 THR LEU LYS MET ASN SER LEU ARG ALA GLU ASP THR ALA SEQRES 8 B 225 VAL TYR TYR CYS ALA LYS LEU GLY THR GLY SER ARG PHE SEQRES 9 B 225 ALA ILE TRP GLY GLN GLY THR LEU VAL THR VAL SER SER SEQRES 10 B 225 ALA SER THR LYS GLY PRO SER VAL PHE PRO LEU ALA PRO SEQRES 11 B 225 SER SER LYS SER THR SER GLY GLY THR ALA ALA LEU GLY SEQRES 12 B 225 CYS LEU VAL LYS ASP TYR PHE PRO GLU PRO VAL THR VAL SEQRES 13 B 225 SER TRP ASN SER GLY ALA LEU THR SER GLY VAL HIS THR SEQRES 14 B 225 PHE PRO ALA VAL LEU GLN SER SER GLY LEU TYR SER LEU SEQRES 15 B 225 SER SER VAL VAL THR VAL PRO SER SER SER LEU GLY THR SEQRES 16 B 225 GLN THR TYR ILE CYS ASN VAL ASN HIS LYS PRO SER ASN SEQRES 17 B 225 THR LYS VAL ASP LYS LYS VAL GLU PRO LYS SER CYS ASP SEQRES 18 B 225 LYS THR HIS THR SEQRES 1 A 218 ASP ILE GLN MET THR GLN SER PRO SER SER LEU SER ALA SEQRES 2 A 218 SER VAL GLY ASP ARG VAL THR ILE THR CYS ARG ALA SER SEQRES 3 A 218 GLN SER VAL TRP ARG ASN LYS PHE VAL ALA TRP TYR GLN SEQRES 4 A 218 GLN LYS PRO GLY LYS ALA PRO LYS ARG LEU ILE TYR ALA SEQRES 5 A 218 ILE ALA SER LEU TYR SER GLY VAL PRO SER ARG PHE SER SEQRES 6 A 218 GLY SER GLY SER GLY THR ASP PHE THR LEU THR ILE SER SEQRES 7 A 218 SER LEU GLN PRO GLU ASP PHE ALA THR TYR TYR CYS VAL SEQRES 8 A 218 GLY HIS TYR GLY SER GLU ASN ASP ALA TYR TYR ALA PHE SEQRES 9 A 218 GLY GLN GLY THR LYS VAL GLU ILE LYS ARG THR VAL ALA SEQRES 10 A 218 ALA PRO SER VAL PHE ILE PHE PRO PRO SER ASP GLU GLN SEQRES 11 A 218 LEU LYS SER GLY THR ALA SER VAL VAL CYS LEU LEU ASN SEQRES 12 A 218 ASN PHE TYR PRO ARG GLU ALA LYS VAL SER TRP TYR VAL SEQRES 13 A 218 ASP ASN ALA LEU GLN SER GLY ASN SER GLN GLU SER VAL SEQRES 14 A 218 THR GLU GLN ASP SER LYS ASP SER THR TYR SER LEU SER SEQRES 15 A 218 SER THR LEU THR LEU SER LYS ALA ASP TYR GLU LYS HIS SEQRES 16 A 218 LYS VAL TYR ALA CYS GLU VAL THR GLN GLY THR THR SER SEQRES 17 A 218 VAL THR LYS SER PHE ASN ARG GLY GLU CYS SEQRES 1 D 9 ALA GLY ALA GLY UKD ALA GLY ALA GLY SEQRES 1 C 9 ALA GLY ALA GLY UKD ALA GLY ALA GLY HET UKD D 5 14 HET UKD C 5 14 HET EDO H 300 10 HET EDO L 301 10 HET EDO B 300 10 HET ACT A 300 7 HETNAM UKD 3-(4-PHOSPHONO-1H-1,2,3-TRIAZOL-1-YL)-L-ALANINE HETNAM EDO 1,2-ETHANEDIOL HETNAM ACT ACETATE ION HETSYN UKD PHIS HETSYN EDO ETHYLENE GLYCOL FORMUL 5 UKD 2(C5 H9 N4 O5 P) FORMUL 7 EDO 3(C2 H6 O2) FORMUL 10 ACT C2 H3 O2 1- FORMUL 11 HOH *188(H2 O) HELIX 1 AA1 SER H 28 TYR H 32 5 5 HELIX 2 AA2 ALA H 60 SER H 65 1 6 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 LYS L 126 1 6 HELIX 10 AB1 LYS L 183 LYS L 188 1 6 HELIX 11 AB2 ARG B 83 THR B 87 5 5 HELIX 12 AB3 SER B 156 ALA B 158 5 3 HELIX 13 AB4 SER B 187 LEU B 189 5 3 HELIX 14 AB5 LYS B 201 ASN B 204 5 4 HELIX 15 AB6 VAL A 27B LYS A 31 5 5 HELIX 16 AB7 GLN A 79 PHE A 83 5 5 HELIX 17 AB8 SER A 121 SER A 127 1 7 HELIX 18 AB9 LYS A 183 GLU A 187 1 5 SHEET 1 AA1 4 GLN H 3 GLY H 8 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 TYR H 90 O THR H 107 SHEET 4 AA2 6 PHE H 34 GLN H 39 -1 N VAL H 37 O TYR H 91 SHEET 5 AA2 6 LEU H 45 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 TYR H 90 O THR H 107 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 LEU L 73 N ILE L 21 SHEET 4 AA7 4 PHE L 62 SER L 67 -1 N SER L 63 O THR L 74 SHEET 1 AA8 6 SER L 10 ALA L 13 0 SHEET 2 AA8 6 THR L 102 ILE L 106 1 O GLU L 105 N LEU L 11 SHEET 3 AA8 6 THR L 85 GLY L 93 -1 N TYR L 86 O THR L 102 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 LYS L 45 N GLN L 37 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 ALA L 13 0 SHEET 2 AA9 4 THR L 102 ILE L 106 1 O GLU L 105 N LEU L 11 SHEET 3 AA9 4 THR L 85 GLY L 93 -1 N TYR L 86 O THR L 102 SHEET 4 AA9 4 ALA L 95C 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 TYR L 173 N PHE L 139 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 B 3 GLY B 8 0 SHEET 2 AB3 4 LEU B 18 SER B 25 -1 O SER B 21 N SER B 7 SHEET 3 AB3 4 THR B 77 MET B 82 -1 O MET B 82 N LEU B 18 SHEET 4 AB3 4 SER B 67 ASN B 72 -1 N THR B 70 O THR B 79 SHEET 1 AB4 6 GLY B 10 VAL B 12 0 SHEET 2 AB4 6 THR B 107 VAL B 111 1 O THR B 110 N GLY B 10 SHEET 3 AB4 6 ALA B 88 LYS B 94 -1 N TYR B 90 O THR B 107 SHEET 4 AB4 6 PHE B 34 GLN B 39 -1 N VAL B 37 O TYR B 91 SHEET 5 AB4 6 LEU B 45 LEU B 51 -1 O ILE B 48 N TRP B 36 SHEET 6 AB4 6 ALA B 57 TYR B 59 -1 O PHE B 58 N TYR B 50 SHEET 1 AB5 4 GLY B 10 VAL B 12 0 SHEET 2 AB5 4 THR B 107 VAL B 111 1 O THR B 110 N GLY B 10 SHEET 3 AB5 4 ALA B 88 LYS B 94 -1 N TYR B 90 O THR B 107 SHEET 4 AB5 4 ILE B 102 TRP B 103 -1 O ILE B 102 N LYS B 94 SHEET 1 AB6 4 SER B 120 LEU B 124 0 SHEET 2 AB6 4 THR B 135 TYR B 145 -1 O LEU B 141 N PHE B 122 SHEET 3 AB6 4 TYR B 176 PRO B 185 -1 O LEU B 178 N VAL B 142 SHEET 4 AB6 4 VAL B 163 THR B 165 -1 N HIS B 164 O VAL B 181 SHEET 1 AB7 4 SER B 120 LEU B 124 0 SHEET 2 AB7 4 THR B 135 TYR B 145 -1 O LEU B 141 N PHE B 122 SHEET 3 AB7 4 TYR B 176 PRO B 185 -1 O LEU B 178 N VAL B 142 SHEET 4 AB7 4 VAL B 169 LEU B 170 -1 N VAL B 169 O SER B 177 SHEET 1 AB8 3 THR B 151 TRP B 154 0 SHEET 2 AB8 3 TYR B 194 HIS B 200 -1 O ASN B 197 N SER B 153 SHEET 3 AB8 3 THR B 205 VAL B 211 -1 O THR B 205 N HIS B 200 SHEET 1 AB9 4 MET A 4 SER A 7 0 SHEET 2 AB9 4 VAL A 19 ALA A 25 -1 O THR A 22 N SER A 7 SHEET 3 AB9 4 ASP A 70 ILE A 75 -1 O LEU A 73 N ILE A 21 SHEET 4 AB9 4 PHE A 62 SER A 67 -1 N SER A 63 O THR A 74 SHEET 1 AC1 6 SER A 10 SER A 14 0 SHEET 2 AC1 6 THR A 102 LYS A 107 1 O LYS A 107 N ALA A 13 SHEET 3 AC1 6 ALA A 84 TYR A 92 -1 N TYR A 86 O THR A 102 SHEET 4 AC1 6 VAL A 33 GLN A 38 -1 N TYR A 36 O TYR A 87 SHEET 5 AC1 6 LYS A 45 TYR A 49 -1 O LYS A 45 N GLN A 37 SHEET 6 AC1 6 SER A 53 LEU A 54 -1 O SER A 53 N TYR A 49 SHEET 1 AC2 4 SER A 10 SER A 14 0 SHEET 2 AC2 4 THR A 102 LYS A 107 1 O LYS A 107 N ALA A 13 SHEET 3 AC2 4 ALA A 84 TYR A 92 -1 N TYR A 86 O THR A 102 SHEET 4 AC2 4 TYR A 95D PHE A 98 -1 O TYR A 95D N TYR A 92 SHEET 1 AC3 4 SER A 114 PHE A 118 0 SHEET 2 AC3 4 THR A 129 PHE A 139 -1 O LEU A 135 N PHE A 116 SHEET 3 AC3 4 TYR A 173 SER A 182 -1 O LEU A 179 N VAL A 132 SHEET 4 AC3 4 SER A 159 VAL A 163 -1 N GLN A 160 O THR A 178 SHEET 1 AC4 4 ALA A 153 LEU A 154 0 SHEET 2 AC4 4 LYS A 145 VAL A 150 -1 N VAL A 150 O ALA A 153 SHEET 3 AC4 4 VAL A 191 THR A 197 -1 O GLU A 195 N SER A 147 SHEET 4 AC4 4 SER A 202 ASN A 208 -1 O VAL A 203 N VAL A 196 SSBOND 1 CYS H 22 CYS H 92 1555 1555 2.01 SSBOND 2 CYS H 140 CYS H 196 1555 1555 2.03 SSBOND 3 CYS H 216 CYS L 212 1555 1555 2.04 SSBOND 4 CYS L 23 CYS L 88 1555 1555 2.11 SSBOND 5 CYS L 134 CYS L 194 1555 1555 2.05 SSBOND 6 CYS B 22 CYS B 92 1555 1555 2.03 SSBOND 7 CYS B 140 CYS B 196 1555 1555 2.00 SSBOND 8 CYS A 23 CYS A 88 1555 1555 2.03 SSBOND 9 CYS A 134 CYS A 194 1555 1555 2.06 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 LINK C GLY C 4 N UKD C 5 1555 1555 1.34 LINK C UKD C 5 N ALA C 6 1555 1555 1.33 CISPEP 1 PHE H 146 PRO H 147 0 -6.42 CISPEP 2 GLU H 148 PRO H 149 0 1.94 CISPEP 3 SER L 7 PRO L 8 0 -8.36 CISPEP 4 TYR L 140 PRO L 141 0 4.68 CISPEP 5 PHE B 146 PRO B 147 0 -8.47 CISPEP 6 GLU B 148 PRO B 149 0 1.27 CISPEP 7 SER A 7 PRO A 8 0 -4.48 CISPEP 8 TYR A 140 PRO A 141 0 7.50 CRYST1 71.546 73.953 239.648 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 0.013977 0.000000 0.000000 0.00000 SCALE2 0.000000 0.013522 0.000000 0.00000 SCALE3 0.000000 0.000000 0.004173 0.00000