HEADER IMMUNE SYSTEM 17-JUN-24 9FQO TITLE CRYSTAL STRUCTURE OF C0083 FAB TARGETING THE OXIDIZED MACROPHAGE TITLE 2 MIGRATION INHIBITORY FACTOR (OXMIF) COMPND MOL_ID: 1; COMPND 2 MOLECULE: C0083 FAB HEAVY CHAIN; COMPND 3 CHAIN: A; COMPND 4 ENGINEERED: YES; COMPND 5 MOL_ID: 2; COMPND 6 MOLECULE: C0083 FAB LIGHT CHAIN; COMPND 7 CHAIN: B; COMPND 8 ENGINEERED: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 3 ORGANISM_TAXID: 9606; SOURCE 4 EXPRESSION_SYSTEM: CRICETULUS GRISEUS; SOURCE 5 EXPRESSION_SYSTEM_TAXID: 10029; SOURCE 6 EXPRESSION_SYSTEM_CELL_LINE: EXPICHO-S; SOURCE 7 EXPRESSION_SYSTEM_ORGAN: OVARY; SOURCE 8 EXPRESSION_SYSTEM_CELL: EPITHELIAL; SOURCE 9 EXPRESSION_SYSTEM_VECTOR_TYPE: PLASMID; SOURCE 10 MOL_ID: 2; SOURCE 11 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 12 ORGANISM_TAXID: 9606; SOURCE 13 EXPRESSION_SYSTEM: CRICETULUS GRISEUS; SOURCE 14 EXPRESSION_SYSTEM_TAXID: 10029; SOURCE 15 EXPRESSION_SYSTEM_CELL_LINE: EXPICHO-S; SOURCE 16 EXPRESSION_SYSTEM_ORGAN: OVARY; SOURCE 17 EXPRESSION_SYSTEM_CELL: EPITHELIAL; SOURCE 18 EXPRESSION_SYSTEM_VECTOR_TYPE: PLASMID KEYWDS FAB, ANTIBODY FRAGMENT, MIF, ANTI-OXMIF, CANCER TREATMENT, KEYWDS 2 IMMUNOTHERAPY, IMMUNE SYSTEM EXPDTA X-RAY DIFFRACTION AUTHOR G.ROSSMUELLER,I.MIRKINA,M.THIELE,A.A.PUCHOL TARAZONA,F.RUEKER, AUTHOR 2 R.J.KERSCHBAUMER,A.SCHINAGL REVDAT 1 01-JAN-25 9FQO 0 JRNL AUTH G.ROSSMUELLER,I.MIRKINA,M.THIELE,A.A.PUCHOL TARAZONA, JRNL AUTH 2 F.RUEKER,R.J.KERSCHBAUMER,A.SCHINAGL JRNL TITL INTEGRATING IN SILICO AND IN VITRO TOOLS FOR OPTIMIZED JRNL TITL 2 ANTIBODY DEVELOPMENT - DESIGN OF THERAPEUTIC ANTI-OXMIF JRNL TITL 3 ANTIBODIES JRNL REF TO BE PUBLISHED JRNL REFN REMARK 1 REMARK 1 REFERENCE 1 REMARK 1 AUTH G.ROSSMUELLER,I.MIRKINA,B.MAURER,V.HOELD,J.MAYER,M.THIELE, REMARK 1 AUTH 2 R.J.KERSCHBAUMER,A.SCHINAGL REMARK 1 TITL PRECLINICAL EVALUATION OF ON203, A NOVEL BIOENGINEERED MAB REMARK 1 TITL 2 TARGETING OXIDIZED MACROPHAGE MIGRATION INHIBITORY FACTOR AS REMARK 1 TITL 3 AN ANTICANCER THERAPEUTIC. REMARK 1 REF MOL CANCER THER V. 22 555 2023 REMARK 1 REFN ESSN 1538-8514 REMARK 1 PMID 37067909 REMARK 1 DOI 10.1158/1535-7163.MCT-22-0676 REMARK 1 REFERENCE 2 REMARK 1 TITL THE NEWLY ENGINEERED MONOCLONAL ANTIBODY ON104, TARGETING REMARK 1 TITL 2 THE OXIDIZED MACROPHAGE MIGRATION INHIBITORY FACTOR (OXMIF), REMARK 1 TITL 3 AMELIORATES CLINICAL AND HISTOPATHOLOGICAL SIGNS OF REMARK 1 TITL 4 COLLAGEN-INDUCED ARTHRITIS. REMARK 1 REF EUR J PHARMACOL V. 956 75997 2023 REMARK 1 REFN ISSN 1879-0712 REMARK 1 PMID 37579967 REMARK 1 DOI 10.1016/J.EJPHAR.2023.175997 REMARK 1 REFERENCE 3 REMARK 1 AUTH A.A.PUCHOL TARAZONA,A.SCHINAGL,I.MIRKINA,G.ROSSMUELLER, REMARK 1 AUTH 2 R.J.KERSCHBAUMER,F.BACHMANN,M.THIELE REMARK 1 TITL PRETARGETED RADIOIMMUNOTHERAPY WITH THE NOVEL ANTI-OXMIF/HSG REMARK 1 TITL 2 BISPECIFIC ANTIBODY ON105 RESULTS IN SIGNIFICANT TUMOR REMARK 1 TITL 3 REGRESSION IN MURINE MODELS OF CANCER. REMARK 1 REF MOL CANCER THER 2024 REMARK 1 REFN ESSN 1538-8514 REMARK 1 PMID 38833646 REMARK 1 DOI 10.1158/1535-7163.MCT-24-0083 REMARK 2 REMARK 2 RESOLUTION. 1.70 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : PHENIX 1.20.1_4487 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) : 1.70 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 41.93 REMARK 3 MIN(FOBS/SIGMA_FOBS) : 1.340 REMARK 3 COMPLETENESS FOR RANGE (%) : 99.7 REMARK 3 NUMBER OF REFLECTIONS : 42855 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 R VALUE (WORKING + TEST SET) : 0.215 REMARK 3 R VALUE (WORKING SET) : 0.215 REMARK 3 FREE R VALUE : 0.237 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 2.170 REMARK 3 FREE R VALUE TEST SET COUNT : 929 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 41.9300 - 3.2500 1.00 6063 135 0.1675 0.1804 REMARK 3 2 3.2500 - 2.5800 1.00 6016 134 0.2125 0.2254 REMARK 3 3 2.5800 - 2.2500 1.00 5983 133 0.2408 0.2607 REMARK 3 4 2.2500 - 2.0500 1.00 5981 133 0.2570 0.3022 REMARK 3 5 2.0500 - 1.9000 1.00 5936 132 0.2927 0.3503 REMARK 3 6 1.9000 - 1.7900 1.00 5940 130 0.3169 0.3663 REMARK 3 7 1.7900 - 1.7000 1.00 5959 133 0.3544 0.3982 REMARK 3 REMARK 3 BULK SOLVENT MODELLING. REMARK 3 METHOD USED : FLAT BULK SOLVENT MODEL REMARK 3 SOLVENT RADIUS : NULL REMARK 3 SHRINKAGE RADIUS : NULL REMARK 3 K_SOL : NULL REMARK 3 B_SOL : NULL REMARK 3 REMARK 3 ERROR ESTIMATES. REMARK 3 COORDINATE ERROR (MAXIMUM-LIKELIHOOD BASED) : NULL REMARK 3 PHASE ERROR (DEGREES, MAXIMUM-LIKELIHOOD BASED) : NULL REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : 27.55 REMARK 3 MEAN B VALUE (OVERALL, A**2) : 37.60 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 3435 REMARK 3 ANGLE : 0.512 4678 REMARK 3 CHIRALITY : 0.043 523 REMARK 3 PLANARITY : 0.004 604 REMARK 3 DIHEDRAL : 4.972 493 REMARK 3 REMARK 3 TLS DETAILS REMARK 3 NUMBER OF TLS GROUPS : 2 REMARK 3 TLS GROUP : 1 REMARK 3 SELECTION: (CHAIN 'A' AND RESID 1 THROUGH 222) REMARK 3 ORIGIN FOR THE GROUP (A): -7.2736 -17.3652 -9.4263 REMARK 3 T TENSOR REMARK 3 T11: 0.2213 T22: 0.3419 REMARK 3 T33: 0.1848 T12: -0.0323 REMARK 3 T13: 0.0396 T23: 0.0070 REMARK 3 L TENSOR REMARK 3 L11: 0.7289 L22: 0.4446 REMARK 3 L33: 2.3476 L12: 0.0896 REMARK 3 L13: 1.2380 L23: 0.2894 REMARK 3 S TENSOR REMARK 3 S11: 0.0735 S12: -0.1607 S13: 0.0247 REMARK 3 S21: 0.0301 S22: -0.0975 S23: -0.0144 REMARK 3 S31: 0.1845 S32: -0.2404 S33: 0.0199 REMARK 3 TLS GROUP : 2 REMARK 3 SELECTION: (CHAIN 'B' AND RESID 1 THROUGH 214) REMARK 3 ORIGIN FOR THE GROUP (A): 0.3505 -6.1369 -21.2569 REMARK 3 T TENSOR REMARK 3 T11: 0.2133 T22: 0.2288 REMARK 3 T33: 0.1677 T12: -0.0134 REMARK 3 T13: 0.0236 T23: -0.0111 REMARK 3 L TENSOR REMARK 3 L11: 1.7848 L22: 0.2371 REMARK 3 L33: 2.8980 L12: 0.2619 REMARK 3 L13: 2.0090 L23: 0.2677 REMARK 3 S TENSOR REMARK 3 S11: -0.0430 S12: 0.1893 S13: -0.0612 REMARK 3 S21: -0.0250 S22: 0.0541 S23: -0.0100 REMARK 3 S31: -0.0966 S32: 0.2772 S33: 0.0015 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 9FQO COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY PDBE ON 17-JUN-24. REMARK 100 THE DEPOSITION ID IS D_1292139184. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 28-APR-23 REMARK 200 TEMPERATURE (KELVIN) : 100 REMARK 200 PH : 6.33 REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : Y REMARK 200 RADIATION SOURCE : ESRF REMARK 200 BEAMLINE : ID30B REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 0.8731 REMARK 200 MONOCHROMATOR : SI(111) REMARK 200 OPTICS : NULL REMARK 200 REMARK 200 DETECTOR TYPE : PIXEL REMARK 200 DETECTOR MANUFACTURER : DECTRIS EIGER2 X 9M REMARK 200 INTENSITY-INTEGRATION SOFTWARE : XDS REMARK 200 DATA SCALING SOFTWARE : XDS REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 42855 REMARK 200 RESOLUTION RANGE HIGH (A) : 1.136 REMARK 200 RESOLUTION RANGE LOW (A) : 50.000 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : NULL REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 99.7 REMARK 200 DATA REDUNDANCY : 6.800 REMARK 200 R MERGE (I) : 0.17500 REMARK 200 R SYM (I) : NULL REMARK 200 FOR THE DATA SET : 9.2200 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 1.70 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 1.76 REMARK 200 COMPLETENESS FOR SHELL (%) : 99.5 REMARK 200 DATA REDUNDANCY IN SHELL : 6.90 REMARK 200 R MERGE FOR SHELL (I) : 1.64700 REMARK 200 R SYM FOR SHELL (I) : NULL REMARK 200 FOR SHELL : 1.440 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 (%): 41.01 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.09 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 0.1M BIS-TRIS, 25.545% (W/V) PEG 3350 REMARK 280 0.2M MGCL2, PH 6.33, VAPOR DIFFUSION, SITTING DROP, TEMPERATURE REMARK 280 293.15K REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 1 21 1 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 -X,Y+1/2,-Z REMARK 290 REMARK 290 WHERE NNN -> OPERATOR NUMBER REMARK 290 MMM -> TRANSLATION VECTOR REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY TRANSFORMATIONS REMARK 290 THE FOLLOWING TRANSFORMATIONS OPERATE ON THE ATOM/HETATM REMARK 290 RECORDS IN THIS ENTRY TO PRODUCE CRYSTALLOGRAPHICALLY REMARK 290 RELATED MOLECULES. REMARK 290 SMTRY1 1 1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 1 0.000000 1.000000 0.000000 0.00000 REMARK 290 SMTRY3 1 0.000000 0.000000 1.000000 0.00000 REMARK 290 SMTRY1 2 -1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 2 0.000000 1.000000 0.000000 33.18000 REMARK 290 SMTRY3 2 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: DIMERIC REMARK 350 APPLY THE FOLLOWING TO CHAINS: A, B 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 LYS A 223 REMARK 465 THR A 224 REMARK 465 HIS A 225 REMARK 465 THR A 226 REMARK 480 REMARK 480 ZERO OCCUPANCY ATOM REMARK 480 THE FOLLOWING RESIDUES HAVE ATOMS MODELED WITH ZERO REMARK 480 OCCUPANCY. THE LOCATION AND PROPERTIES OF THESE ATOMS REMARK 480 MAY NOT BE RELIABLE. (M=MODEL NUMBER; RES=RESIDUE NAME; REMARK 480 C=CHAIN IDENTIFIER; SSEQ=SEQUENCE NUMBER; I=INSERTION CODE): REMARK 480 M RES C SSEQI ATOMS REMARK 480 CYS B 134 N CA C O CB SG 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 O HOH B 308 O HOH B 413 2.05 REMARK 500 O HOH B 348 O HOH B 367 2.06 REMARK 500 O HOH A 480 O HOH B 365 2.09 REMARK 500 O HOH A 510 O HOH A 515 2.12 REMARK 500 REMARK 500 REMARK: NULL REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: CLOSE CONTACTS REMARK 500 REMARK 500 THE FOLLOWING ATOMS THAT ARE RELATED BY CRYSTALLOGRAPHIC REMARK 500 SYMMETRY ARE IN CLOSE CONTACT. AN ATOM LOCATED WITHIN 0.15 REMARK 500 ANGSTROMS OF A SYMMETRY RELATED ATOM IS ASSUMED TO BE ON A REMARK 500 SPECIAL POSITION AND IS, THEREFORE, LISTED IN REMARK 375 REMARK 500 INSTEAD OF REMARK 500. ATOMS WITH NON-BLANK ALTERNATE REMARK 500 LOCATION INDICATORS ARE NOT INCLUDED IN THE CALCULATIONS. REMARK 500 REMARK 500 DISTANCE CUTOFF: REMARK 500 2.2 ANGSTROMS FOR CONTACTS NOT INVOLVING HYDROGEN ATOMS REMARK 500 1.6 ANGSTROMS FOR CONTACTS INVOLVING HYDROGEN ATOMS REMARK 500 REMARK 500 ATM1 RES C SSEQI ATM2 RES C SSEQI SSYMOP DISTANCE REMARK 500 NH2 ARG A 87 OG SER B 203 1556 2.08 REMARK 500 OG1 THR A 196 O ARG B 18 2544 2.15 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 SER A 7 166.00 177.12 REMARK 500 ALA A 92 170.67 176.83 REMARK 500 THR A 136 47.78 -99.53 REMARK 500 ASP A 149 73.60 64.25 REMARK 500 THR A 196 -46.70 -134.61 REMARK 500 MET B 30 -117.15 53.60 REMARK 500 LEU B 47 -60.46 -104.83 REMARK 500 GLU B 68 -95.68 59.22 REMARK 500 ALA B 84 -179.78 -174.53 REMARK 500 PHE B 92 -66.03 -90.76 REMARK 500 ASN B 138 70.57 59.96 REMARK 500 REMARK 500 REMARK: NULL REMARK 525 REMARK 525 SOLVENT REMARK 525 REMARK 525 THE SOLVENT MOLECULES HAVE CHAIN IDENTIFIERS THAT REMARK 525 INDICATE THE POLYMER CHAIN WITH WHICH THEY ARE MOST REMARK 525 CLOSELY ASSOCIATED. THE REMARK LISTS ALL THE SOLVENT REMARK 525 MOLECULES WHICH ARE MORE THAN 5A AWAY FROM THE REMARK 525 NEAREST POLYMER CHAIN (M = MODEL NUMBER; REMARK 525 RES=RESIDUE NAME; C=CHAIN IDENTIFIER; SSEQ=SEQUENCE REMARK 525 NUMBER; I=INSERTION CODE): REMARK 525 REMARK 525 M RES CSSEQI REMARK 525 HOH A 520 DISTANCE = 6.93 ANGSTROMS REMARK 525 HOH A 521 DISTANCE = 6.95 ANGSTROMS DBREF 9FQO A 1 226 PDB 9FQO 9FQO 1 226 DBREF 9FQO B 1 214 PDB 9FQO 9FQO 1 214 SEQRES 1 A 226 GLU VAL GLN LEU GLN GLU SER GLY GLY GLY LEU VAL GLN SEQRES 2 A 226 PRO GLY GLY SER LEU ARG LEU SER CYS ALA ALA SER GLY SEQRES 3 A 226 PHE THR PHE SER ILE TYR SER MET ASN TRP VAL ARG GLN SEQRES 4 A 226 ALA PRO GLY LYS GLY LEU GLU TRP VAL SER SER ILE GLY SEQRES 5 A 226 SER SER GLY GLY THR THR TYR TYR ALA ASP SER VAL LYS SEQRES 6 A 226 GLY ARG PHE THR ILE SER ARG ASP ASN SER LYS ASN THR SEQRES 7 A 226 LEU TYR LEU GLN MET ASN SER LEU ARG ALA GLU ASP THR SEQRES 8 A 226 ALA VAL TYR TYR CYS ALA GLY SER GLN TYR LEU TYR GLY SEQRES 9 A 226 MET ASP VAL TRP GLY GLN GLY THR THR VAL THR VAL SER SEQRES 10 A 226 SER ALA SER THR LYS GLY PRO SER VAL PHE PRO LEU ALA SEQRES 11 A 226 PRO SER SER LYS SER THR SER GLY GLY THR ALA ALA LEU SEQRES 12 A 226 GLY CYS LEU VAL LYS ASP TYR PHE PRO GLU PRO VAL THR SEQRES 13 A 226 VAL SER TRP ASN SER GLY ALA LEU THR SER GLY VAL HIS SEQRES 14 A 226 THR PHE PRO ALA VAL LEU GLN SER SER GLY LEU TYR SER SEQRES 15 A 226 LEU SER SER VAL VAL THR VAL PRO SER SER SER LEU GLY SEQRES 16 A 226 THR GLN THR TYR ILE CYS ASN VAL ASN HIS LYS PRO SER SEQRES 17 A 226 ASN THR LYS VAL ASP LYS ARG VAL GLU PRO LYS SER CYS SEQRES 18 A 226 ASP LYS THR HIS THR SEQRES 1 B 214 ASP ILE GLN MET THR GLN SER PRO SER SER LEU SER ALA SEQRES 2 B 214 SER VAL GLY ASP ARG VAL THR ILE THR CYS ARG SER SER SEQRES 3 B 214 GLN ARG ILE MET THR TYR LEU ASN TRP TYR GLN GLN LYS SEQRES 4 B 214 PRO GLY LYS ALA PRO LYS LEU LEU ILE TYR VAL GLY SER SEQRES 5 B 214 HIS SER GLN SER GLY VAL PRO SER ARG PHE ARG GLY SER SEQRES 6 B 214 GLY SER GLU THR ASP PHE THR LEU THR ILE SER GLY LEU SEQRES 7 B 214 GLN PRO GLU ASP SER ALA THR TYR TYR CYS GLN GLN SER SEQRES 8 B 214 PHE PHE THR PRO LEU THR PHE GLY GLY GLY THR LYS VAL SEQRES 9 B 214 GLU ILE LYS ARG THR VAL ALA ALA PRO SER VAL PHE ILE SEQRES 10 B 214 PHE PRO PRO SER ASP GLU GLN LEU LYS SER GLY THR ALA SEQRES 11 B 214 SER VAL VAL CYS LEU LEU ASN ASN PHE TYR PRO ARG GLU SEQRES 12 B 214 ALA LYS VAL GLN TRP LYS VAL ASP ASN ALA LEU GLN SER SEQRES 13 B 214 GLY ASN SER GLN GLU SER VAL THR GLU GLN ASP SER LYS SEQRES 14 B 214 ASP SER THR TYR SER LEU SER SER THR LEU THR LEU SER SEQRES 15 B 214 LYS ALA ASP TYR GLU LYS HIS LYS VAL TYR ALA CYS GLU SEQRES 16 B 214 VAL THR HIS GLN GLY LEU SER SER PRO VAL THR LYS SER SEQRES 17 B 214 PHE ASN ARG GLY GLU CYS HET BTB A 301 33 HETNAM BTB 2-[BIS-(2-HYDROXY-ETHYL)-AMINO]-2-HYDROXYMETHYL- HETNAM 2 BTB PROPANE-1,3-DIOL HETSYN BTB BIS-TRIS BUFFER FORMUL 3 BTB C8 H19 N O5 FORMUL 4 HOH *260(H2 O) HELIX 1 AA1 THR A 28 TYR A 32 5 5 HELIX 2 AA2 ARG A 87 THR A 91 5 5 HELIX 3 AA3 SER A 161 ALA A 163 5 3 HELIX 4 AA4 SER A 192 LEU A 194 5 3 HELIX 5 AA5 GLN B 79 SER B 83 5 5 HELIX 6 AA6 SER B 121 SER B 127 1 7 HELIX 7 AA7 LYS B 183 GLU B 187 1 5 SHEET 1 AA1 4 GLN A 3 SER A 7 0 SHEET 2 AA1 4 LEU A 18 SER A 25 -1 O ALA A 23 N GLN A 5 SHEET 3 AA1 4 THR A 78 MET A 83 -1 O MET A 83 N LEU A 18 SHEET 4 AA1 4 PHE A 68 ASP A 73 -1 N SER A 71 O TYR A 80 SHEET 1 AA2 6 GLY A 10 VAL A 12 0 SHEET 2 AA2 6 THR A 112 VAL A 116 1 O THR A 115 N VAL A 12 SHEET 3 AA2 6 ALA A 92 SER A 99 -1 N TYR A 94 O THR A 112 SHEET 4 AA2 6 SER A 33 GLN A 39 -1 N VAL A 37 O TYR A 95 SHEET 5 AA2 6 LEU A 45 ILE A 51 -1 O GLU A 46 N ARG A 38 SHEET 6 AA2 6 THR A 58 TYR A 60 -1 O TYR A 59 N SER A 50 SHEET 1 AA3 4 GLY A 10 VAL A 12 0 SHEET 2 AA3 4 THR A 112 VAL A 116 1 O THR A 115 N VAL A 12 SHEET 3 AA3 4 ALA A 92 SER A 99 -1 N TYR A 94 O THR A 112 SHEET 4 AA3 4 VAL A 107 TRP A 108 -1 N VAL A 107 O GLY A 98 SHEET 1 AA4 4 SER A 125 LEU A 129 0 SHEET 2 AA4 4 THR A 140 TYR A 150 -1 O LEU A 146 N PHE A 127 SHEET 3 AA4 4 TYR A 181 PRO A 190 -1 O VAL A 189 N ALA A 141 SHEET 4 AA4 4 VAL A 168 THR A 170 -1 N HIS A 169 O VAL A 186 SHEET 1 AA5 4 SER A 125 LEU A 129 0 SHEET 2 AA5 4 THR A 140 TYR A 150 -1 O LEU A 146 N PHE A 127 SHEET 3 AA5 4 TYR A 181 PRO A 190 -1 O VAL A 189 N ALA A 141 SHEET 4 AA5 4 VAL A 174 LEU A 175 -1 N VAL A 174 O SER A 182 SHEET 1 AA6 3 THR A 156 TRP A 159 0 SHEET 2 AA6 3 ILE A 200 HIS A 205 -1 O ASN A 202 N SER A 158 SHEET 3 AA6 3 THR A 210 ARG A 215 -1 O VAL A 212 N VAL A 203 SHEET 1 AA7 4 MET B 4 SER B 7 0 SHEET 2 AA7 4 ARG B 18 SER B 25 -1 O ARG B 24 N THR B 5 SHEET 3 AA7 4 ASP B 70 SER B 76 -1 O LEU B 73 N ILE B 21 SHEET 4 AA7 4 PHE B 62 SER B 67 -1 N SER B 65 O THR B 72 SHEET 1 AA8 6 SER B 10 SER B 14 0 SHEET 2 AA8 6 THR B 102 LYS B 107 1 O GLU B 105 N LEU B 11 SHEET 3 AA8 6 ALA B 84 GLN B 90 -1 N ALA B 84 O VAL B 104 SHEET 4 AA8 6 LEU B 33 GLN B 38 -1 N ASN B 34 O GLN B 89 SHEET 5 AA8 6 LYS B 45 TYR B 49 -1 O LEU B 47 N TRP B 35 SHEET 6 AA8 6 HIS B 53 SER B 54 -1 O HIS B 53 N TYR B 49 SHEET 1 AA9 4 SER B 10 SER B 14 0 SHEET 2 AA9 4 THR B 102 LYS B 107 1 O GLU B 105 N LEU B 11 SHEET 3 AA9 4 ALA B 84 GLN B 90 -1 N ALA B 84 O VAL B 104 SHEET 4 AA9 4 THR B 97 PHE B 98 -1 O THR B 97 N GLN B 90 SHEET 1 AB1 4 SER B 114 PHE B 118 0 SHEET 2 AB1 4 THR B 129 PHE B 139 -1 O LEU B 135 N PHE B 116 SHEET 3 AB1 4 TYR B 173 SER B 182 -1 O LEU B 179 N VAL B 132 SHEET 4 AB1 4 SER B 159 VAL B 163 -1 N SER B 162 O SER B 176 SHEET 1 AB2 4 ALA B 153 LEU B 154 0 SHEET 2 AB2 4 LYS B 145 VAL B 150 -1 N VAL B 150 O ALA B 153 SHEET 3 AB2 4 VAL B 191 THR B 197 -1 O GLU B 195 N GLN B 147 SHEET 4 AB2 4 VAL B 205 ASN B 210 -1 O VAL B 205 N VAL B 196 SSBOND 1 CYS A 22 CYS A 96 1555 1555 2.04 SSBOND 2 CYS A 145 CYS A 201 1555 1555 2.04 SSBOND 3 CYS B 23 CYS B 88 1555 1555 2.04 SSBOND 4 CYS B 134 CYS B 194 1555 1555 2.03 CISPEP 1 PHE A 151 PRO A 152 0 -2.68 CISPEP 2 GLU A 153 PRO A 154 0 0.21 CISPEP 3 SER B 7 PRO B 8 0 -0.57 CISPEP 4 THR B 94 PRO B 95 0 1.45 CISPEP 5 TYR B 140 PRO B 141 0 1.40 CRYST1 51.540 66.360 58.650 90.00 100.08 90.00 P 1 21 1 2 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 0.019402 0.000000 0.003449 0.00000 SCALE2 0.000000 0.015069 0.000000 0.00000 SCALE3 0.000000 0.000000 0.017318 0.00000