HEADER IMMUNE SYSTEM 09-OCT-23 8UHS TITLE ANTI-PHOSPHOHISTIDINE FAB HSC44.CK.20.ELBOW BOUND TO PHOSPHATE COMPND MOL_ID: 1; COMPND 2 MOLECULE: HSC44.CK.20.ELBOW FAB HEAVY CHAIN; COMPND 3 CHAIN: H; COMPND 4 ENGINEERED: YES; COMPND 5 OTHER_DETAILS: FAB HEAVY CHAIN; COMPND 6 MOL_ID: 2; COMPND 7 MOLECULE: HSC44.CK.20.ELBOW FAB LIGHT CHAIN; COMPND 8 CHAIN: L; COMPND 9 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 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 8UHS 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. 2.40 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) : 2.40 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 49.79 REMARK 3 MIN(FOBS/SIGMA_FOBS) : 1.330 REMARK 3 COMPLETENESS FOR RANGE (%) : 99.8 REMARK 3 NUMBER OF REFLECTIONS : 22300 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 R VALUE (WORKING + TEST SET) : 0.253 REMARK 3 R VALUE (WORKING SET) : 0.252 REMARK 3 FREE R VALUE : 0.286 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 4.840 REMARK 3 FREE R VALUE TEST SET COUNT : 1079 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 49.7900 - 4.7900 1.00 2875 142 0.1976 0.2378 REMARK 3 2 4.7900 - 3.8100 1.00 2691 138 0.2073 0.2189 REMARK 3 3 3.8000 - 3.3200 1.00 2654 138 0.2805 0.3246 REMARK 3 4 3.3200 - 3.0200 1.00 2613 137 0.2934 0.3308 REMARK 3 5 3.0200 - 2.8000 1.00 2649 122 0.3085 0.3645 REMARK 3 6 2.8000 - 2.6400 1.00 2617 119 0.3459 0.3781 REMARK 3 7 2.6400 - 2.5100 1.00 2573 149 0.3279 0.3704 REMARK 3 8 2.5100 - 2.4000 0.99 2549 134 0.3614 0.4370 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) : 0.439 REMARK 3 PHASE ERROR (DEGREES, MAXIMUM-LIKELIHOOD BASED) : 30.928 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : 51.14 REMARK 3 MEAN B VALUE (OVERALL, A**2) : 75.75 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 : 0.002 3368 REMARK 3 ANGLE : 0.535 4576 REMARK 3 CHIRALITY : 0.042 515 REMARK 3 PLANARITY : 0.005 583 REMARK 3 DIHEDRAL : 12.813 1195 REMARK 3 REMARK 3 TLS DETAILS REMARK 3 NUMBER OF TLS GROUPS : 5 REMARK 3 TLS GROUP : 1 REMARK 3 SELECTION: CHAIN 'H' AND (RESID 1 THROUGH 106 ) REMARK 3 ORIGIN FOR THE GROUP (A): -3.3260 -35.3171 4.4215 REMARK 3 T TENSOR REMARK 3 T11: 0.5203 T22: 1.0055 REMARK 3 T33: 0.6715 T12: 0.1239 REMARK 3 T13: 0.0728 T23: 0.4229 REMARK 3 L TENSOR REMARK 3 L11: 1.7584 L22: 2.7232 REMARK 3 L33: 0.4206 L12: 0.9047 REMARK 3 L13: -0.4236 L23: -0.4112 REMARK 3 S TENSOR REMARK 3 S11: 0.0225 S12: 0.1270 S13: -0.4092 REMARK 3 S21: 0.3686 S22: 1.1608 S23: 0.4055 REMARK 3 S31: -0.1979 S32: -1.3914 S33: -0.3660 REMARK 3 TLS GROUP : 2 REMARK 3 SELECTION: CHAIN 'H' AND (RESID 107 THROUGH 133 ) REMARK 3 ORIGIN FOR THE GROUP (A): 14.2484 -27.3299 -22.2333 REMARK 3 T TENSOR REMARK 3 T11: 0.4373 T22: 0.3458 REMARK 3 T33: 0.3713 T12: -0.0173 REMARK 3 T13: -0.0114 T23: 0.0719 REMARK 3 L TENSOR REMARK 3 L11: 1.5061 L22: 0.7035 REMARK 3 L33: 2.7079 L12: -0.6623 REMARK 3 L13: 0.5353 L23: 0.7880 REMARK 3 S TENSOR REMARK 3 S11: -0.1960 S12: 0.0416 S13: 0.0914 REMARK 3 S21: -0.3138 S22: 0.1981 S23: 0.0603 REMARK 3 S31: 0.0009 S32: -0.1621 S33: -0.0216 REMARK 3 TLS GROUP : 3 REMARK 3 SELECTION: CHAIN 'H' AND (RESID 134 THROUGH 213 ) REMARK 3 ORIGIN FOR THE GROUP (A): 19.9407 -25.3141 -20.4369 REMARK 3 T TENSOR REMARK 3 T11: 0.5112 T22: 0.2628 REMARK 3 T33: 0.3688 T12: -0.0212 REMARK 3 T13: -0.0222 T23: -0.0503 REMARK 3 L TENSOR REMARK 3 L11: 1.6532 L22: 2.3518 REMARK 3 L33: 1.3810 L12: -0.1541 REMARK 3 L13: -0.1033 L23: -0.2619 REMARK 3 S TENSOR REMARK 3 S11: -0.3029 S12: -0.0937 S13: -0.0417 REMARK 3 S21: 0.3582 S22: 0.2056 S23: -0.3055 REMARK 3 S31: 0.2015 S32: 0.1042 S33: 0.0647 REMARK 3 TLS GROUP : 4 REMARK 3 SELECTION: CHAIN 'L' AND (RESID 1 THROUGH 113 ) REMARK 3 ORIGIN FOR THE GROUP (A): -0.4375 -13.8068 7.0025 REMARK 3 T TENSOR REMARK 3 T11: 1.6081 T22: 0.7017 REMARK 3 T33: 0.8228 T12: 0.7579 REMARK 3 T13: 0.8236 T23: 0.0698 REMARK 3 L TENSOR REMARK 3 L11: 0.0796 L22: 0.8025 REMARK 3 L33: 1.9001 L12: 0.0229 REMARK 3 L13: -0.0489 L23: -0.5284 REMARK 3 S TENSOR REMARK 3 S11: 0.4357 S12: -0.2500 S13: 0.8181 REMARK 3 S21: 1.4926 S22: 0.7867 S23: 1.5149 REMARK 3 S31: -1.7710 S32: -0.9407 S33: 0.1774 REMARK 3 TLS GROUP : 5 REMARK 3 SELECTION: CHAIN 'L' AND (RESID 114 THROUGH 212 ) REMARK 3 ORIGIN FOR THE GROUP (A): 11.2664 -13.1218 -27.7979 REMARK 3 T TENSOR REMARK 3 T11: 0.4999 T22: 0.2719 REMARK 3 T33: 0.3929 T12: -0.0125 REMARK 3 T13: 0.0033 T23: 0.0390 REMARK 3 L TENSOR REMARK 3 L11: 1.7837 L22: 3.1993 REMARK 3 L33: 2.1764 L12: 0.4470 REMARK 3 L13: 0.2420 L23: -0.3828 REMARK 3 S TENSOR REMARK 3 S11: 0.0012 S12: 0.0310 S13: 0.0746 REMARK 3 S21: 0.0412 S22: 0.3018 S23: 0.2809 REMARK 3 S31: -0.1994 S32: -0.1125 S33: -0.2792 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 8UHS COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 17-OCT-23. REMARK 100 THE DEPOSITION ID IS D_1000277438. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 01-DEC-22 REMARK 200 TEMPERATURE (KELVIN) : 100 REMARK 200 PH : 7.5 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 : 22383 REMARK 200 RESOLUTION RANGE HIGH (A) : 2.400 REMARK 200 RESOLUTION RANGE LOW (A) : 49.790 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : -3.000 REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 100.0 REMARK 200 DATA REDUNDANCY : 9.800 REMARK 200 R MERGE (I) : 0.07100 REMARK 200 R SYM (I) : 0.07100 REMARK 200 FOR THE DATA SET : 42.0000 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 2.40 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 2.44 REMARK 200 COMPLETENESS FOR SHELL (%) : 99.7 REMARK 200 DATA REDUNDANCY IN SHELL : 6.20 REMARK 200 R MERGE FOR SHELL (I) : 0.76100 REMARK 200 R SYM FOR SHELL (I) : 0.76100 REMARK 200 FOR SHELL : 2.480 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 (%): 56.57 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.83 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 0.1M HEPES, PH 7.5, 20% PEG4000, 10% 2 REMARK 280 -PROPANOL, VAPOR DIFFUSION, SITTING DROP, TEMPERATURE 293K REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 43 21 2 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 -X,-Y,Z+1/2 REMARK 290 3555 -Y+1/2,X+1/2,Z+3/4 REMARK 290 4555 Y+1/2,-X+1/2,Z+1/4 REMARK 290 5555 -X+1/2,Y+1/2,-Z+3/4 REMARK 290 6555 X+1/2,-Y+1/2,-Z+1/4 REMARK 290 7555 Y,X,-Z REMARK 290 8555 -Y,-X,-Z+1/2 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 102.96500 REMARK 290 SMTRY1 3 0.000000 -1.000000 0.000000 36.16300 REMARK 290 SMTRY2 3 1.000000 0.000000 0.000000 36.16300 REMARK 290 SMTRY3 3 0.000000 0.000000 1.000000 154.44750 REMARK 290 SMTRY1 4 0.000000 1.000000 0.000000 36.16300 REMARK 290 SMTRY2 4 -1.000000 0.000000 0.000000 36.16300 REMARK 290 SMTRY3 4 0.000000 0.000000 1.000000 51.48250 REMARK 290 SMTRY1 5 -1.000000 0.000000 0.000000 36.16300 REMARK 290 SMTRY2 5 0.000000 1.000000 0.000000 36.16300 REMARK 290 SMTRY3 5 0.000000 0.000000 -1.000000 154.44750 REMARK 290 SMTRY1 6 1.000000 0.000000 0.000000 36.16300 REMARK 290 SMTRY2 6 0.000000 -1.000000 0.000000 36.16300 REMARK 290 SMTRY3 6 0.000000 0.000000 -1.000000 51.48250 REMARK 290 SMTRY1 7 0.000000 1.000000 0.000000 0.00000 REMARK 290 SMTRY2 7 1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY3 7 0.000000 0.000000 -1.000000 0.00000 REMARK 290 SMTRY1 8 0.000000 -1.000000 0.000000 0.00000 REMARK 290 SMTRY2 8 -1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY3 8 0.000000 0.000000 -1.000000 102.96500 REMARK 290 REMARK 290 REMARK: NULL REMARK 300 REMARK 300 BIOMOLECULE: 1 REMARK 300 SEE REMARK 350 FOR THE AUTHOR PROVIDED AND/OR PROGRAM REMARK 300 GENERATED ASSEMBLY INFORMATION FOR THE STRUCTURE IN REMARK 300 THIS ENTRY. THE REMARK MAY ALSO PROVIDE INFORMATION ON REMARK 300 BURIED SURFACE AREA. REMARK 350 REMARK 350 COORDINATES FOR A COMPLETE MULTIMER REPRESENTING THE KNOWN REMARK 350 BIOLOGICALLY SIGNIFICANT OLIGOMERIZATION STATE OF THE REMARK 350 MOLECULE CAN BE GENERATED BY APPLYING BIOMT TRANSFORMATIONS REMARK 350 GIVEN BELOW. BOTH NON-CRYSTALLOGRAPHIC AND REMARK 350 CRYSTALLOGRAPHIC OPERATIONS ARE GIVEN. REMARK 350 REMARK 350 BIOMOLECULE: 1 REMARK 350 AUTHOR DETERMINED BIOLOGICAL UNIT: DIMERIC REMARK 350 SOFTWARE DETERMINED QUATERNARY STRUCTURE: DIMERIC REMARK 350 SOFTWARE USED: PISA REMARK 350 TOTAL BURIED SURFACE AREA: 4380 ANGSTROM**2 REMARK 350 SURFACE AREA OF THE COMPLEX: 19660 ANGSTROM**2 REMARK 350 CHANGE IN SOLVENT FREE ENERGY: -26.0 KCAL/MOL REMARK 350 APPLY THE FOLLOWING TO CHAINS: H, L 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 SER H 126 REMARK 465 SER H 214 REMARK 465 CYS H 215 REMARK 465 ASP H 216 REMARK 465 LYS H 217 REMARK 465 THR H 218 REMARK 465 HIS H 219 REMARK 465 THR H 220 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 GLN L 27 -168.93 -123.22 REMARK 500 ILE L 51 -51.19 64.98 REMARK 500 SER L 94 -158.55 -150.23 REMARK 500 ASN L 138 74.38 55.52 REMARK 500 REMARK 500 REMARK: NULL DBREF 8UHS H 1 220 PDB 8UHS 8UHS 1 220 DBREF 8UHS L 1 212 PDB 8UHS 8UHS 1 212 SEQRES 1 H 224 GLU VAL GLN LEU VAL GLU SER GLY GLY GLY LEU VAL GLN SEQRES 2 H 224 PRO GLY GLY SER LEU ARG LEU SER CYS THR ALA SER GLY SEQRES 3 H 224 PHE SER ILE ASP SER TYR GLY PHE SER TRP VAL ARG GLN SEQRES 4 H 224 ALA PRO GLY LYS GLY LEU GLU HIS ILE GLY TYR LEU THR SEQRES 5 H 224 ALA GLY GLY ARG ALA PHE TYR ALA SER TRP ALA LYS SER SEQRES 6 H 224 ARG SER THR ILE THR ARG ASN THR ASN GLU ASN THR VAL SEQRES 7 H 224 THR LEU LYS MET ASN SER LEU ARG ALA GLU ASP THR ALA SEQRES 8 H 224 VAL TYR TYR CYS ALA LYS LEU GLY THR GLY SER ARG PHE SEQRES 9 H 224 ALA ILE TRP GLY GLN GLY THR LEU VAL THR VAL PHE ASN SEQRES 10 H 224 GLN ILE LYS GLY PRO SER VAL PHE PRO LEU ALA PRO SER SEQRES 11 H 224 SER LYS SER THR SER GLY GLY THR ALA ALA LEU GLY CYS SEQRES 12 H 224 LEU VAL LYS ASP TYR PHE PRO GLU PRO VAL THR VAL SER SEQRES 13 H 224 TRP ASN SER GLY ALA LEU THR SER GLY VAL HIS THR PHE SEQRES 14 H 224 PRO ALA VAL LEU GLN SER SER GLY LEU TYR SER LEU SER SEQRES 15 H 224 SER VAL VAL THR VAL PRO SER SER SER LEU GLY THR GLN SEQRES 16 H 224 THR TYR ILE CYS ASN VAL ASN HIS LYS PRO SER ASN THR SEQRES 17 H 224 LYS VAL ASP LYS LYS VAL GLU PRO LYS SER CYS ASP LYS SEQRES 18 H 224 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 HET EDO H 301 4 HET PO4 H 302 5 HETNAM EDO 1,2-ETHANEDIOL HETNAM PO4 PHOSPHATE ION HETSYN EDO ETHYLENE GLYCOL FORMUL 3 EDO C2 H6 O2 FORMUL 4 PO4 O4 P 3- FORMUL 5 HOH *24(H2 O) HELIX 1 AA1 ALA H 60 SER H 65 1 6 HELIX 2 AA2 ARG H 83 THR H 87 5 5 HELIX 3 AA3 SER H 155 ALA H 157 5 3 HELIX 4 AA4 SER H 186 LEU H 188 5 3 HELIX 5 AA5 LYS H 200 ASN H 203 5 4 HELIX 6 AA6 GLN L 79 PHE L 83 5 5 HELIX 7 AA7 SER L 121 LYS L 126 1 6 HELIX 8 AA8 LYS L 183 GLU L 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 THR H 23 N VAL H 5 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 ASN H 72 O THR H 77 SHEET 1 AA2 6 LEU H 11 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 SER H 35 O ALA H 93 SHEET 5 AA2 6 LEU H 45 LEU H 51 -1 O LEU H 51 N PHE H 34 SHEET 6 AA2 6 ALA H 57 TYR H 59 -1 O PHE H 58 N TYR H 50 SHEET 1 AA3 4 LEU H 11 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 119 LEU H 123 0 SHEET 2 AA4 4 THR H 134 TYR H 144 -1 O LEU H 140 N PHE H 121 SHEET 3 AA4 4 TYR H 175 PRO H 184 -1 O LEU H 177 N VAL H 141 SHEET 4 AA4 4 HIS H 163 THR H 164 -1 N HIS H 163 O VAL H 180 SHEET 1 AA5 4 SER H 119 LEU H 123 0 SHEET 2 AA5 4 THR H 134 TYR H 144 -1 O LEU H 140 N PHE H 121 SHEET 3 AA5 4 TYR H 175 PRO H 184 -1 O LEU H 177 N VAL H 141 SHEET 4 AA5 4 VAL H 168 LEU H 169 -1 N VAL H 168 O SER H 176 SHEET 1 AA6 3 THR H 150 TRP H 153 0 SHEET 2 AA6 3 ILE H 194 HIS H 199 -1 O ASN H 196 N SER H 152 SHEET 3 AA6 3 THR H 204 LYS H 209 -1 O VAL H 206 N VAL H 197 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 GLY L 64 -1 N SER L 63 O THR L 74 SHEET 1 AA8 2 SER L 10 ALA L 13 0 SHEET 2 AA8 2 LYS L 103 ILE L 106 1 O LYS L 103 N LEU L 11 SHEET 1 AA9 5 SER L 53 LEU L 54 0 SHEET 2 AA9 5 LYS L 45 TYR L 49 -1 N TYR L 49 O SER L 53 SHEET 3 AA9 5 VAL L 33 GLN L 38 -1 N GLN L 37 O LYS L 45 SHEET 4 AA9 5 THR L 85 HIS L 91 -1 O TYR L 87 N TYR L 36 SHEET 5 AA9 5 TYR L 96 PHE L 98 -1 O ALA L 97 N GLY L 90 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 181 N ALA L 130 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 LYS L 205 N CYS L 194 SSBOND 1 CYS H 22 CYS H 92 1555 1555 2.03 SSBOND 2 CYS H 139 CYS H 195 1555 1555 2.03 SSBOND 3 CYS L 23 CYS L 88 1555 1555 2.04 SSBOND 4 CYS L 134 CYS L 194 1555 1555 2.04 CISPEP 1 PHE H 145 PRO H 146 0 -3.41 CISPEP 2 GLU H 147 PRO H 148 0 -2.55 CISPEP 3 SER L 7 PRO L 8 0 -5.40 CISPEP 4 TYR L 140 PRO L 141 0 3.19 CRYST1 72.326 72.326 205.930 90.00 90.00 90.00 P 43 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 0.013826 0.000000 0.000000 0.00000 SCALE2 0.000000 0.013826 0.000000 0.00000 SCALE3 0.000000 0.000000 0.004856 0.00000