HEADER SIGNALING PROTEIN 21-OCT-22 8BE4 TITLE CRYSTAL STRUCTURE OF SOS1-KRASG12V-NANOBODY14 COMPND MOL_ID: 1; COMPND 2 MOLECULE: SON OF SEVENLESS HOMOLOG 1; COMPND 3 CHAIN: S; COMPND 4 SYNONYM: SOS-1; COMPND 5 ENGINEERED: YES; COMPND 6 MOL_ID: 2; COMPND 7 MOLECULE: ISOFORM 2B OF GTPASE KRAS; COMPND 8 CHAIN: R; COMPND 9 SYNONYM: K-RAS 2,KI-RAS,C-K-RAS,C-KI-RAS; COMPND 10 EC: 3.6.5.2; COMPND 11 ENGINEERED: YES; COMPND 12 MUTATION: YES; COMPND 13 MOL_ID: 3; COMPND 14 MOLECULE: NANOBODY14; COMPND 15 CHAIN: C000; COMPND 16 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: SOS1; SOURCE 6 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 7 EXPRESSION_SYSTEM_TAXID: 562; SOURCE 8 MOL_ID: 2; SOURCE 9 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 10 ORGANISM_COMMON: HUMAN; SOURCE 11 ORGANISM_TAXID: 9606; SOURCE 12 GENE: KRAS, KRAS2, RASK2; SOURCE 13 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 14 EXPRESSION_SYSTEM_TAXID: 562; SOURCE 15 MOL_ID: 3; SOURCE 16 ORGANISM_SCIENTIFIC: LAMA GLAMA; SOURCE 17 ORGANISM_TAXID: 9844; SOURCE 18 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 19 EXPRESSION_SYSTEM_TAXID: 562 KEYWDS COMPLEX, STABILIZER, SIGNALING PROTEIN EXPDTA X-RAY DIFFRACTION AUTHOR B.FISCHER,A.WOHLKONIG,J.STEYAERT JRNL AUTH B.FISCHER,A.WOHLKONIG,J.STEYAERT JRNL TITL CHILL & DISCO TO DISCOVER NANOBODIES THAT MODULATE JRNL TITL 2 PROTEIN-PROTEIN INTERACTIONS AND TUNE THE SOS-RAS NUCLEOTIDE JRNL TITL 3 EXCHANGE RATE JRNL REF TO BE PUBLISHED JRNL REFN REMARK 2 REMARK 2 RESOLUTION. 1.90 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) : 1.90 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 29.59 REMARK 3 MIN(FOBS/SIGMA_FOBS) : 1.370 REMARK 3 COMPLETENESS FOR RANGE (%) : 99.6 REMARK 3 NUMBER OF REFLECTIONS : 189895 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 R VALUE (WORKING + TEST SET) : 0.176 REMARK 3 R VALUE (WORKING SET) : 0.175 REMARK 3 FREE R VALUE : 0.199 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 5.010 REMARK 3 FREE R VALUE TEST SET COUNT : 9506 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 29.5900 - 5.8700 1.00 6041 313 0.1603 0.1600 REMARK 3 2 5.8700 - 4.6700 1.00 6048 321 0.1496 0.1462 REMARK 3 3 4.6700 - 4.0800 1.00 5985 319 0.1302 0.1405 REMARK 3 4 4.0800 - 3.7100 1.00 6077 319 0.1463 0.1764 REMARK 3 5 3.7100 - 3.4400 1.00 6038 319 0.1648 0.1885 REMARK 3 6 3.4400 - 3.2400 1.00 6080 315 0.1785 0.2358 REMARK 3 7 3.2400 - 3.0800 1.00 6012 319 0.1862 0.2048 REMARK 3 8 3.0800 - 2.9400 1.00 5991 319 0.1853 0.1960 REMARK 3 9 2.9400 - 2.8300 1.00 6056 319 0.1809 0.2242 REMARK 3 10 2.8300 - 2.7300 1.00 6047 320 0.1829 0.2483 REMARK 3 11 2.7300 - 2.6500 1.00 6004 318 0.1962 0.2499 REMARK 3 12 2.6500 - 2.5700 1.00 6068 317 0.1936 0.2250 REMARK 3 13 2.5700 - 2.5000 1.00 6021 319 0.1898 0.2454 REMARK 3 14 2.5000 - 2.4400 1.00 6061 320 0.1833 0.1860 REMARK 3 15 2.4400 - 2.3900 1.00 6054 321 0.1778 0.2146 REMARK 3 16 2.3900 - 2.3400 1.00 6028 316 0.1804 0.1926 REMARK 3 17 2.3400 - 2.2900 1.00 5994 315 0.1844 0.2086 REMARK 3 18 2.2900 - 2.2500 1.00 6058 320 0.1886 0.2212 REMARK 3 19 2.2500 - 2.2100 1.00 6028 318 0.1875 0.1948 REMARK 3 20 2.2100 - 2.1700 1.00 6063 319 0.1945 0.2452 REMARK 3 21 2.1700 - 2.1300 1.00 5996 317 0.1963 0.2350 REMARK 3 22 2.1300 - 2.1000 1.00 6046 319 0.2068 0.2688 REMARK 3 23 2.1000 - 2.0700 1.00 5983 314 0.2343 0.2490 REMARK 3 24 2.0700 - 2.0400 1.00 6134 326 0.2322 0.2578 REMARK 3 25 2.0400 - 2.0100 1.00 6050 318 0.2328 0.2583 REMARK 3 26 2.0100 - 1.9900 1.00 6037 318 0.2400 0.2739 REMARK 3 27 1.9900 - 1.9600 1.00 5946 316 0.2468 0.2772 REMARK 3 28 1.9600 - 1.9400 1.00 6055 318 0.2572 0.2906 REMARK 3 29 1.9400 - 1.9200 1.00 6039 317 0.2826 0.3121 REMARK 3 30 1.9200 - 1.9000 0.88 5349 277 0.3483 0.4002 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) : 0.229 REMARK 3 PHASE ERROR (DEGREES, MAXIMUM-LIKELIHOOD BASED) : 21.128 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : 35.62 REMARK 3 MEAN B VALUE (OVERALL, A**2) : 41.67 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 : 0.019 6215 REMARK 3 ANGLE : 1.385 8421 REMARK 3 CHIRALITY : 0.104 916 REMARK 3 PLANARITY : 0.009 1100 REMARK 3 DIHEDRAL : 15.479 2387 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 8BE4 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_1292126144. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 30-JUL-17 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 : DIAMOND REMARK 200 BEAMLINE : I03 REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 0.97622 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 : XDS REMARK 200 DATA SCALING SOFTWARE : XDS REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 189905 REMARK 200 RESOLUTION RANGE HIGH (A) : 1.895 REMARK 200 RESOLUTION RANGE LOW (A) : 29.590 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : NULL REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 99.6 REMARK 200 DATA REDUNDANCY : 6.900 REMARK 200 R MERGE (I) : NULL REMARK 200 R SYM (I) : NULL REMARK 200 FOR THE DATA SET : 15.6300 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 1.90 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 2.01 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: PHENIX REMARK 200 STARTING MODEL: 1BKD REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 62.26 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 3.26 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 14% PEG 1000, TRIS 0.1 M PH 7.0, VAPOR REMARK 280 DIFFUSION, SITTING DROP, TEMPERATURE 293K REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: I 4 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 -Y,X,Z REMARK 290 4555 Y,-X,Z REMARK 290 5555 X+1/2,Y+1/2,Z+1/2 REMARK 290 6555 -X+1/2,-Y+1/2,Z+1/2 REMARK 290 7555 -Y+1/2,X+1/2,Z+1/2 REMARK 290 8555 Y+1/2,-X+1/2,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 0.00000 REMARK 290 SMTRY1 3 0.000000 -1.000000 0.000000 0.00000 REMARK 290 SMTRY2 3 1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY3 3 0.000000 0.000000 1.000000 0.00000 REMARK 290 SMTRY1 4 0.000000 1.000000 0.000000 0.00000 REMARK 290 SMTRY2 4 -1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY3 4 0.000000 0.000000 1.000000 0.00000 REMARK 290 SMTRY1 5 1.000000 0.000000 0.000000 63.83750 REMARK 290 SMTRY2 5 0.000000 1.000000 0.000000 63.83750 REMARK 290 SMTRY3 5 0.000000 0.000000 1.000000 76.04650 REMARK 290 SMTRY1 6 -1.000000 0.000000 0.000000 63.83750 REMARK 290 SMTRY2 6 0.000000 -1.000000 0.000000 63.83750 REMARK 290 SMTRY3 6 0.000000 0.000000 1.000000 76.04650 REMARK 290 SMTRY1 7 0.000000 -1.000000 0.000000 63.83750 REMARK 290 SMTRY2 7 1.000000 0.000000 0.000000 63.83750 REMARK 290 SMTRY3 7 0.000000 0.000000 1.000000 76.04650 REMARK 290 SMTRY1 8 0.000000 1.000000 0.000000 63.83750 REMARK 290 SMTRY2 8 -1.000000 0.000000 0.000000 63.83750 REMARK 290 SMTRY3 8 0.000000 0.000000 1.000000 76.04650 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: TRIMERIC REMARK 350 APPLY THE FOLLOWING TO CHAINS: S, R, C000 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 MET S 543 REMARK 465 GLY S 544 REMARK 465 SER S 545 REMARK 465 SER S 546 REMARK 465 HIS S 547 REMARK 465 HIS S 548 REMARK 465 HIS S 549 REMARK 465 HIS S 550 REMARK 465 HIS S 551 REMARK 465 HIS S 552 REMARK 465 SER S 553 REMARK 465 SER S 554 REMARK 465 GLY S 555 REMARK 465 LEU S 556 REMARK 465 VAL S 557 REMARK 465 PRO S 558 REMARK 465 ARG S 559 REMARK 465 GLY S 560 REMARK 465 SER S 561 REMARK 465 HIS S 562 REMARK 465 MET S 563 REMARK 465 GLU S 564 REMARK 465 GLU S 565 REMARK 465 GLN S 566 REMARK 465 MET S 567 REMARK 465 ARG S 568 REMARK 465 LEU S 569 REMARK 465 MET S 592 REMARK 465 GLN S 593 REMARK 465 PRO S 594 REMARK 465 LYS S 595 REMARK 465 ALA S 596 REMARK 465 PRO S 653 REMARK 465 GLU S 654 REMARK 465 PRO S 655 REMARK 465 THR S 656 REMARK 465 GLU S 657 REMARK 465 ALA S 658 REMARK 465 ASP S 659 REMARK 465 ARG S 660 REMARK 465 ILE S 661 REMARK 465 ALA S 662 REMARK 465 ILE S 663 REMARK 465 GLU S 664 REMARK 465 ASN S 665 REMARK 465 GLY S 666 REMARK 465 ASP S 667 REMARK 465 GLN S 668 REMARK 465 PRO S 669 REMARK 465 LEU S 670 REMARK 465 SER S 671 REMARK 465 ALA S 672 REMARK 465 ARG S 744 REMARK 465 ASP S 745 REMARK 465 ASN S 746 REMARK 465 GLY S 747 REMARK 465 PRO S 748 REMARK 465 GLY S 749 REMARK 465 HIS S 750 REMARK 465 ASN S 751 REMARK 465 ILE S 752 REMARK 465 THR S 753 REMARK 465 PRO S 1045 REMARK 465 ARG S 1046 REMARK 465 PRO S 1047 REMARK 465 GLY S 1048 REMARK 465 THR S 1049 REMARK 465 MET R -20 REMARK 465 GLY R -19 REMARK 465 SER R -18 REMARK 465 SER R -17 REMARK 465 HIS R -16 REMARK 465 HIS R -15 REMARK 465 HIS R -14 REMARK 465 HIS R -13 REMARK 465 HIS R -12 REMARK 465 HIS R -11 REMARK 465 SER R -10 REMARK 465 SER R -9 REMARK 465 GLY R -8 REMARK 465 GLU R -7 REMARK 465 ASN R -6 REMARK 465 LEU R -5 REMARK 465 TYR R -4 REMARK 465 PHE R -3 REMARK 465 GLN R -2 REMARK 465 GLY R -1 REMARK 465 SER R 0 REMARK 465 CYS R 118 REMARK 465 ASP R 119 REMARK 465 LEU R 120 REMARK 465 LYS R 169 REMARK 465 HIS C 120 REMARK 465 HIS C 121 REMARK 465 HIS C 122 REMARK 465 HIS C 123 REMARK 465 HIS C 124 REMARK 465 GLU C 125 REMARK 465 PRO C 126 REMARK 465 GLU C 127 REMARK 465 ALA C 128 REMARK 470 REMARK 470 MISSING ATOM REMARK 470 THE FOLLOWING RESIDUES HAVE MISSING ATOMS (M=MODEL NUMBER; REMARK 470 RES=RESIDUE NAME; C=CHAIN IDENTIFIER; SSEQ=SEQUENCE NUMBER; REMARK 470 I=INSERTION CODE): REMARK 470 M RES CSSEQI ATOMS REMARK 470 ARG S 722 CG CD NE CZ NH1 NH2 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 HIS S 764 -87.61 -118.16 REMARK 500 ALA00 91 172.78 179.34 REMARK 500 REMARK 500 REMARK: NULL DBREF 8BE4 S 564 1049 UNP Q07889 SOS1_HUMAN 564 1049 DBREF 8BE4 R 1 169 UNP P01116-2 RASK_HUMAN 1 169 DBREF 8BE400 1 128 PDB 8BE4 8BE4 1 128 SEQADV 8BE4 MET S 543 UNP Q07889 INITIATING METHIONINE SEQADV 8BE4 GLY S 544 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 SER S 545 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 SER S 546 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 HIS S 547 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 HIS S 548 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 HIS S 549 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 HIS S 550 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 HIS S 551 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 HIS S 552 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 SER S 553 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 SER S 554 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 GLY S 555 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 LEU S 556 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 VAL S 557 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 PRO S 558 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 ARG S 559 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 GLY S 560 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 SER S 561 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 HIS S 562 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 MET S 563 UNP Q07889 EXPRESSION TAG SEQADV 8BE4 MET R -20 UNP P01116-2 INITIATING METHIONINE SEQADV 8BE4 GLY R -19 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 SER R -18 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 SER R -17 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 HIS R -16 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 HIS R -15 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 HIS R -14 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 HIS R -13 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 HIS R -12 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 HIS R -11 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 SER R -10 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 SER R -9 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 GLY R -8 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 GLU R -7 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 ASN R -6 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 LEU R -5 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 TYR R -4 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 PHE R -3 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 GLN R -2 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 GLY R -1 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 SER R 0 UNP P01116-2 EXPRESSION TAG SEQADV 8BE4 VAL R 12 UNP P01116-2 GLY 12 ENGINEERED MUTATION SEQRES 1 S 507 MET GLY SER SER HIS HIS HIS HIS HIS HIS SER SER GLY SEQRES 2 S 507 LEU VAL PRO ARG GLY SER HIS MET GLU GLU GLN MET ARG SEQRES 3 S 507 LEU PRO SER ALA ASP VAL TYR ARG PHE ALA GLU PRO ASP SEQRES 4 S 507 SER GLU GLU ASN ILE ILE PHE GLU GLU ASN MET GLN PRO SEQRES 5 S 507 LYS ALA GLY ILE PRO ILE ILE LYS ALA GLY THR VAL ILE SEQRES 6 S 507 LYS LEU ILE GLU ARG LEU THR TYR HIS MET TYR ALA ASP SEQRES 7 S 507 PRO ASN PHE VAL ARG THR PHE LEU THR THR TYR ARG SER SEQRES 8 S 507 PHE CYS LYS PRO GLN GLU LEU LEU SER LEU ILE ILE GLU SEQRES 9 S 507 ARG PHE GLU ILE PRO GLU PRO GLU PRO THR GLU ALA ASP SEQRES 10 S 507 ARG ILE ALA ILE GLU ASN GLY ASP GLN PRO LEU SER ALA SEQRES 11 S 507 GLU LEU LYS ARG PHE ARG LYS GLU TYR ILE GLN PRO VAL SEQRES 12 S 507 GLN LEU ARG VAL LEU ASN VAL CYS ARG HIS TRP VAL GLU SEQRES 13 S 507 HIS HIS PHE TYR ASP PHE GLU ARG ASP ALA TYR LEU LEU SEQRES 14 S 507 GLN ARG MET GLU GLU PHE ILE GLY THR VAL ARG GLY LYS SEQRES 15 S 507 ALA MET LYS LYS TRP VAL GLU SER ILE THR LYS ILE ILE SEQRES 16 S 507 GLN ARG LYS LYS ILE ALA ARG ASP ASN GLY PRO GLY HIS SEQRES 17 S 507 ASN ILE THR PHE GLN SER SER PRO PRO THR VAL GLU TRP SEQRES 18 S 507 HIS ILE SER ARG PRO GLY HIS ILE GLU THR PHE ASP LEU SEQRES 19 S 507 LEU THR LEU HIS PRO ILE GLU ILE ALA ARG GLN LEU THR SEQRES 20 S 507 LEU LEU GLU SER ASP LEU TYR ARG ALA VAL GLN PRO SER SEQRES 21 S 507 GLU LEU VAL GLY SER VAL TRP THR LYS GLU ASP LYS GLU SEQRES 22 S 507 ILE ASN SER PRO ASN LEU LEU LYS MET ILE ARG HIS THR SEQRES 23 S 507 THR ASN LEU THR LEU TRP PHE GLU LYS CYS ILE VAL GLU SEQRES 24 S 507 THR GLU ASN LEU GLU GLU ARG VAL ALA VAL VAL SER ARG SEQRES 25 S 507 ILE ILE GLU ILE LEU GLN VAL PHE GLN GLU LEU ASN ASN SEQRES 26 S 507 PHE ASN GLY VAL LEU GLU VAL VAL SER ALA MET ASN SER SEQRES 27 S 507 SER PRO VAL TYR ARG LEU ASP HIS THR PHE GLU GLN ILE SEQRES 28 S 507 PRO SER ARG GLN LYS LYS ILE LEU GLU GLU ALA HIS GLU SEQRES 29 S 507 LEU SER GLU ASP HIS TYR LYS LYS TYR LEU ALA LYS LEU SEQRES 30 S 507 ARG SER ILE ASN PRO PRO CYS VAL PRO PHE PHE GLY ILE SEQRES 31 S 507 TYR LEU THR ASN ILE LEU LYS THR GLU GLU GLY ASN PRO SEQRES 32 S 507 GLU VAL LEU LYS ARG HIS GLY LYS GLU LEU ILE ASN PHE SEQRES 33 S 507 SER LYS ARG ARG LYS VAL ALA GLU ILE THR GLY GLU ILE SEQRES 34 S 507 GLN GLN TYR GLN ASN GLN PRO TYR CYS LEU ARG VAL GLU SEQRES 35 S 507 SER ASP ILE LYS ARG PHE PHE GLU ASN LEU ASN PRO MET SEQRES 36 S 507 GLY ASN SER MET GLU LYS GLU PHE THR ASP TYR LEU PHE SEQRES 37 S 507 ASN LYS SER LEU GLU ILE GLU PRO ARG ASN PRO LYS PRO SEQRES 38 S 507 LEU PRO ARG PHE PRO LYS LYS TYR SER TYR PRO LEU LYS SEQRES 39 S 507 SER PRO GLY VAL ARG PRO SER ASN PRO ARG PRO GLY THR SEQRES 1 R 190 MET GLY SER SER HIS HIS HIS HIS HIS HIS SER SER GLY SEQRES 2 R 190 GLU ASN LEU TYR PHE GLN GLY SER MET THR GLU TYR LYS SEQRES 3 R 190 LEU VAL VAL VAL GLY ALA VAL GLY VAL GLY LYS SER ALA SEQRES 4 R 190 LEU THR ILE GLN LEU ILE GLN ASN HIS PHE VAL ASP GLU SEQRES 5 R 190 TYR ASP PRO THR ILE GLU ASP SER TYR ARG LYS GLN VAL SEQRES 6 R 190 VAL ILE ASP GLY GLU THR CYS LEU LEU ASP ILE LEU ASP SEQRES 7 R 190 THR ALA GLY GLN GLU GLU TYR SER ALA MET ARG ASP GLN SEQRES 8 R 190 TYR MET ARG THR GLY GLU GLY PHE LEU CYS VAL PHE ALA SEQRES 9 R 190 ILE ASN ASN THR LYS SER PHE GLU ASP ILE HIS HIS TYR SEQRES 10 R 190 ARG GLU GLN ILE LYS ARG VAL LYS ASP SER GLU ASP VAL SEQRES 11 R 190 PRO MET VAL LEU VAL GLY ASN LYS CYS ASP LEU PRO SER SEQRES 12 R 190 ARG THR VAL ASP THR LYS GLN ALA GLN ASP LEU ALA ARG SEQRES 13 R 190 SER TYR GLY ILE PRO PHE ILE GLU THR SER ALA LYS THR SEQRES 14 R 190 ARG GLN GLY VAL ASP ASP ALA PHE TYR THR LEU VAL ARG SEQRES 15 R 190 GLU ILE ARG LYS HIS LYS GLU LYS SEQRES 100 128 GLN VAL GLN LEU VAL GLU SER GLY GLY GLY LEU VAL GLN SEQRES 200 128 ALA GLY GLY SER LEU ARG LEU SER CYS ALA ALA SER ARG SEQRES 300 128 SER SER PHE THR ILE ASN ARG MET GLY TRP TYR ARG GLN SEQRES 400 128 ALA PRO GLY LYS GLN ARG GLU LEU VAL ALA ASP ILE THR SEQRES 500 128 SER GLY GLY ASN ARG ASN TYR ALA ASP SER VAL LYS GLY SEQRES 600 128 ARG PHE THR ILE ALA ARG ASP ASN ALA LYS ASN THR ALA SEQRES 700 128 TYR LEU GLN MET ASN SER LEU LYS PRO GLU ASP THR ALA SEQRES 800 128 VAL TYR TYR CYS ASN ALA LYS ILE HIS PRO TRP SER VAL SEQRES 900 128 ALA ASP LEU TRP GLY GLN GLY THR GLN VAL THR VAL SER SEQRES 1000 128 SER HIS HIS HIS HIS HIS HIS GLU PRO GLU ALA FORMUL 4 HOH *551(H2 O) HELIX 1 AA1 TYR S 575 GLU S 579 5 5 HELIX 2 AA2 THR S 605 THR S 614 1 10 HELIX 3 AA3 ASP S 620 TYR S 631 1 12 HELIX 4 AA4 ARG S 632 PHE S 634 5 3 HELIX 5 AA5 LYS S 636 GLU S 649 1 14 HELIX 6 AA6 LEU S 674 TYR S 681 1 8 HELIX 7 AA7 TYR S 681 HIS S 700 1 20 HELIX 8 AA8 PHE S 701 ASP S 707 1 7 HELIX 9 AA9 ASP S 707 THR S 720 1 14 HELIX 10 AB1 MET S 726 ILE S 742 1 17 HELIX 11 AB2 HIS S 770 PHE S 774 5 5 HELIX 12 AB3 HIS S 780 ALA S 798 1 19 HELIX 13 AB4 GLN S 800 LYS S 811 5 12 HELIX 14 AB5 ASP S 813 SER S 818 1 6 HELIX 15 AB6 SER S 818 GLU S 841 1 24 HELIX 16 AB7 ASN S 844 LEU S 865 1 22 HELIX 17 AB8 ASN S 867 SER S 880 1 14 HELIX 18 AB9 SER S 880 ARG S 885 1 6 HELIX 19 AC1 LEU S 886 GLN S 892 1 7 HELIX 20 AC2 PRO S 894 LEU S 907 1 14 HELIX 21 AC3 SER S 908 ILE S 922 1 15 HELIX 22 AC4 PHE S 930 GLY S 943 1 14 HELIX 23 AC5 PHE S 958 TYR S 974 1 17 HELIX 24 AC6 GLU S 984 ASN S 993 1 10 HELIX 25 AC7 MET S 1001 GLU S 1017 1 17 HELIX 26 AC8 GLY R 15 GLN R 25 1 11 HELIX 27 AC9 ILE R 36 SER R 39 5 4 HELIX 28 AD1 TYR R 64 ALA R 66 5 3 HELIX 29 AD2 MET R 67 THR R 74 1 8 HELIX 30 AD3 ASN R 86 ASP R 105 1 20 HELIX 31 AD4 ASP R 126 GLY R 138 1 13 HELIX 32 AD5 GLY R 151 GLU R 168 1 18 HELIX 33 AD6 ASP00 61 LYS00 64 5 4 HELIX 34 AD7 ASN00 73 LYS00 75 5 3 HELIX 35 AD8 LYS00 86 THR00 90 5 5 SHEET 1 AA1 4 ILE S 586 PHE S 588 0 SHEET 2 AA1 4 ILE S 601 GLY S 604 -1 O ALA S 603 N ILE S 587 SHEET 3 AA1 4 LYS S 953 ASN S 957 -1 O ILE S 956 N GLY S 604 SHEET 4 AA1 4 VAL S 947 ARG S 950 -1 N LEU S 948 O LEU S 955 SHEET 1 AA2 6 ARG R 41 ILE R 46 0 SHEET 2 AA2 6 GLU R 49 ASP R 57 -1 O GLU R 49 N ILE R 46 SHEET 3 AA2 6 THR R 2 GLY R 10 1 N LEU R 6 O LEU R 56 SHEET 4 AA2 6 GLY R 77 ALA R 83 1 O VAL R 81 N VAL R 9 SHEET 5 AA2 6 MET R 111 ASN R 116 1 O ASN R 116 N PHE R 82 SHEET 6 AA2 6 PHE R 141 THR R 144 1 O ILE R 142 N LEU R 113 SHEET 1 AA3 4 LEU00 4 SER00 7 0 SHEET 2 AA3 4 LEU00 18 ALA00 24 -1 O SER00 21 N SER00 7 SHEET 3 AA3 4 THR00 77 MET00 82 -1 O MET00 82 N LEU00 18 SHEET 4 AA3 4 PHE00 67 ASP00 72 -1 N THR00 68 O GLN00 81 SHEET 1 AA4 6 GLY00 10 GLN00 13 0 SHEET 2 AA4 6 THR00 112 SER00 117 1 O GLN00 113 N GLY00 10 SHEET 3 AA4 6 ALA00 91 LYS00 98 -1 N TYR00 93 O THR00 112 SHEET 4 AA4 6 ARG00 33 GLN00 39 -1 N TYR00 37 O TYR00 94 SHEET 5 AA4 6 GLU00 46 THR00 52 -1 O GLU00 46 N ARG00 38 SHEET 6 AA4 6 ARG00 57 TYR00 59 -1 O ASN00 58 N ASP00 50 SHEET 1 AA5 4 GLY00 10 GLN00 13 0 SHEET 2 AA5 4 THR00 112 SER00 117 1 O GLN00 113 N GLY00 10 SHEET 3 AA5 4 ALA00 91 LYS00 98 -1 N TYR00 93 O THR00 112 SHEET 4 AA5 4 LEU00 107 TRP00 108 -1 O LEU00 107 N ALA00 97 SSBOND 1 CYS00 22 CYS00 95 1555 1555 2.06 CISPEP 1 PRO S 924 PRO S 925 0 6.91 CISPEP 2 ASN S 1020 PRO S 1021 0 -2.72 CRYST1 127.675 127.675 152.093 90.00 90.00 90.00 I 4 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 0.007832 0.000000 0.000000 0.00000 SCALE2 0.000000 0.007832 0.000000 0.00000 SCALE3 0.000000 0.000000 0.006575 0.00000