HEADER IMMUNE SYSTEM 14-APR-24 8Z39 TITLE CRYSTAL STRUCTURE OF SONIC HEDGEHOG IN COMPLEX WITH ANTIBODY 5E1 TITLE 2 MUTANT L-T56A WITH METALS COMPND MOL_ID: 1; COMPND 2 MOLECULE: SONIC HEDGEHOG PROTEIN N-PRODUCT; COMPND 3 CHAIN: A; COMPND 4 SYNONYM: SHHN,SHH N-TERMINAL PROCESSED SIGNALING DOMAINS,SHHNP; COMPND 5 ENGINEERED: YES; COMPND 6 MOL_ID: 2; COMPND 7 MOLECULE: HEAVY CHAIN OF ANTIBODY 5E1; COMPND 8 CHAIN: H; COMPND 9 ENGINEERED: YES; COMPND 10 MOL_ID: 3; COMPND 11 MOLECULE: LIGHT CHAIN OF ANTIBODY 5E1; COMPND 12 CHAIN: L; COMPND 13 ENGINEERED: YES; COMPND 14 MUTATION: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 3 ORGANISM_COMMON: HUMAN; SOURCE 4 ORGANISM_TAXID: 9606; SOURCE 5 GENE: SHH; SOURCE 6 EXPRESSION_SYSTEM: ESCHERICHIA COLI BL21(DE3); SOURCE 7 EXPRESSION_SYSTEM_TAXID: 469008; SOURCE 8 MOL_ID: 2; SOURCE 9 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 10 ORGANISM_TAXID: 9606; SOURCE 11 GENE: IGG; SOURCE 12 EXPRESSION_SYSTEM: HOMO SAPIENS; SOURCE 13 EXPRESSION_SYSTEM_TAXID: 9606; SOURCE 14 EXPRESSION_SYSTEM_CELL_LINE: EXPI293; SOURCE 15 MOL_ID: 3; SOURCE 16 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 17 ORGANISM_TAXID: 9606; SOURCE 18 GENE: IGG; SOURCE 19 EXPRESSION_SYSTEM: HOMO SAPIENS; SOURCE 20 EXPRESSION_SYSTEM_TAXID: 9606; SOURCE 21 EXPRESSION_SYSTEM_CELL_LINE: EXPI293 KEYWDS SONIC HEDGEHOG PROTEIN (SHH), ANTIBODY, FLEXIBLE/FLUCTUATED EPITOPE, KEYWDS 2 AFFINITY IMPROVEMENT, IMMUNE SYSTEM EXPDTA X-RAY DIFFRACTION AUTHOR J.M.M.CAAVEIRO,I.KANEDA,K.TSUMOTO REVDAT 1 16-APR-25 8Z39 0 JRNL AUTH I.KANEDA,R.MATSUNAGA,S.NAGATOISHI,A.SENOO,J.M.M.CAAVEIRO, JRNL AUTH 2 D.KURODA,K.TSUMOTO JRNL TITL BIOPHYSICAL INSIGHTS TO IMPROVE AFFINITY OF ANTIBODIES TO JRNL TITL 2 FLUCTUATED EPITOPE OF ANTIGEN: A CASE OF ANTI-SHH ANTIBODY JRNL TITL 3 5E1 AGAINST SHH ANTIGEN. JRNL REF TO BE PUBLISHED JRNL REFN REMARK 2 REMARK 2 RESOLUTION. 1.80 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : REFMAC 5.8.0238 REMARK 3 AUTHORS : MURSHUDOV,SKUBAK,LEBEDEV,PANNU,STEINER, REMARK 3 : NICHOLLS,WINN,LONG,VAGIN REMARK 3 REMARK 3 REFINEMENT TARGET : MAXIMUM LIKELIHOOD REMARK 3 REMARK 3 DATA USED IN REFINEMENT. REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 1.80 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 48.65 REMARK 3 DATA CUTOFF (SIGMA(F)) : NULL REMARK 3 COMPLETENESS FOR RANGE (%) : 99.9 REMARK 3 NUMBER OF REFLECTIONS : 67860 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 CROSS-VALIDATION METHOD : THROUGHOUT REMARK 3 FREE R VALUE TEST SET SELECTION : RANDOM REMARK 3 R VALUE (WORKING + TEST SET) : 0.181 REMARK 3 R VALUE (WORKING SET) : 0.180 REMARK 3 FREE R VALUE : 0.208 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 4.000 REMARK 3 FREE R VALUE TEST SET COUNT : 2817 REMARK 3 REMARK 3 FIT IN THE HIGHEST RESOLUTION BIN. REMARK 3 TOTAL NUMBER OF BINS USED : 20 REMARK 3 BIN RESOLUTION RANGE HIGH (A) : 1.80 REMARK 3 BIN RESOLUTION RANGE LOW (A) : 1.85 REMARK 3 REFLECTION IN BIN (WORKING SET) : 4973 REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 99.94 REMARK 3 BIN R VALUE (WORKING SET) : 0.2900 REMARK 3 BIN FREE R VALUE SET COUNT : 208 REMARK 3 BIN FREE R VALUE : 0.2960 REMARK 3 REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT. REMARK 3 PROTEIN ATOMS : 4487 REMARK 3 NUCLEIC ACID ATOMS : 0 REMARK 3 HETEROGEN ATOMS : 9 REMARK 3 SOLVENT ATOMS : 625 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : NULL REMARK 3 MEAN B VALUE (OVERALL, A**2) : 29.71 REMARK 3 OVERALL ANISOTROPIC B VALUE. REMARK 3 B11 (A**2) : 1.56000 REMARK 3 B22 (A**2) : -1.43000 REMARK 3 B33 (A**2) : 0.57000 REMARK 3 B12 (A**2) : 0.00000 REMARK 3 B13 (A**2) : -1.21000 REMARK 3 B23 (A**2) : 0.00000 REMARK 3 REMARK 3 ESTIMATED OVERALL COORDINATE ERROR. REMARK 3 ESU BASED ON R VALUE (A): 0.109 REMARK 3 ESU BASED ON FREE R VALUE (A): 0.105 REMARK 3 ESU BASED ON MAXIMUM LIKELIHOOD (A): 0.099 REMARK 3 ESU FOR B VALUES BASED ON MAXIMUM LIKELIHOOD (A**2): 6.300 REMARK 3 REMARK 3 CORRELATION COEFFICIENTS. REMARK 3 CORRELATION COEFFICIENT FO-FC : 0.967 REMARK 3 CORRELATION COEFFICIENT FO-FC FREE : 0.954 REMARK 3 REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES COUNT RMS WEIGHT REMARK 3 BOND LENGTHS REFINED ATOMS (A): 4698 ; 0.012 ; 0.013 REMARK 3 BOND LENGTHS OTHERS (A): 4222 ; 0.001 ; 0.017 REMARK 3 BOND ANGLES REFINED ATOMS (DEGREES): 6408 ; 1.676 ; 1.646 REMARK 3 BOND ANGLES OTHERS (DEGREES): 9874 ; 1.379 ; 1.576 REMARK 3 TORSION ANGLES, PERIOD 1 (DEGREES): 623 ;17.279 ; 5.249 REMARK 3 TORSION ANGLES, PERIOD 2 (DEGREES): 224 ;32.160 ;23.304 REMARK 3 TORSION ANGLES, PERIOD 3 (DEGREES): 772 ;13.650 ;15.000 REMARK 3 TORSION ANGLES, PERIOD 4 (DEGREES): 21 ;18.751 ;15.000 REMARK 3 CHIRAL-CENTER RESTRAINTS (A**3): 618 ; 0.072 ; 0.200 REMARK 3 GENERAL PLANES REFINED ATOMS (A): 5880 ; 0.009 ; 0.020 REMARK 3 GENERAL PLANES OTHERS (A): 967 ; 0.001 ; 0.020 REMARK 3 NON-BONDED CONTACTS REFINED ATOMS (A): NULL ; NULL ; NULL REMARK 3 NON-BONDED CONTACTS OTHERS (A): NULL ; NULL ; NULL REMARK 3 NON-BONDED TORSION REFINED ATOMS (A): NULL ; NULL ; NULL REMARK 3 NON-BONDED TORSION OTHERS (A): NULL ; NULL ; NULL REMARK 3 H-BOND (X...Y) REFINED ATOMS (A): NULL ; NULL ; NULL REMARK 3 H-BOND (X...Y) OTHERS (A): NULL ; NULL ; NULL REMARK 3 POTENTIAL METAL-ION REFINED ATOMS (A): NULL ; NULL ; NULL REMARK 3 POTENTIAL METAL-ION OTHERS (A): NULL ; NULL ; NULL REMARK 3 SYMMETRY VDW REFINED ATOMS (A): NULL ; NULL ; NULL REMARK 3 SYMMETRY VDW OTHERS (A): NULL ; NULL ; NULL REMARK 3 SYMMETRY H-BOND REFINED ATOMS (A): NULL ; NULL ; NULL REMARK 3 SYMMETRY H-BOND OTHERS (A): NULL ; NULL ; NULL REMARK 3 SYMMETRY METAL-ION REFINED ATOMS (A): NULL ; NULL ; NULL REMARK 3 SYMMETRY METAL-ION OTHERS (A): NULL ; NULL ; NULL REMARK 3 REMARK 3 ISOTROPIC THERMAL FACTOR RESTRAINTS. COUNT RMS WEIGHT REMARK 3 MAIN-CHAIN BOND REFINED ATOMS (A**2): 2342 ; 1.103 ; 1.760 REMARK 3 MAIN-CHAIN BOND OTHER ATOMS (A**2): 2340 ; 1.097 ; 1.757 REMARK 3 MAIN-CHAIN ANGLE REFINED ATOMS (A**2): 2929 ; 1.845 ; 2.631 REMARK 3 MAIN-CHAIN ANGLE OTHER ATOMS (A**2): 2929 ; 1.846 ; 2.630 REMARK 3 SIDE-CHAIN BOND REFINED ATOMS (A**2): 2356 ; 1.403 ; 1.912 REMARK 3 SIDE-CHAIN BOND OTHER ATOMS (A**2): 2353 ; 1.403 ; 1.913 REMARK 3 SIDE-CHAIN ANGLE REFINED ATOMS (A**2): NULL ; NULL ; NULL REMARK 3 SIDE-CHAIN ANGLE OTHER ATOMS (A**2): 3457 ; 2.296 ; 2.796 REMARK 3 LONG RANGE B REFINED ATOMS (A**2): 5340 ; 5.067 ;21.933 REMARK 3 LONG RANGE B OTHER ATOMS (A**2): 5174 ; 4.925 ;20.962 REMARK 3 REMARK 3 ANISOTROPIC THERMAL FACTOR RESTRAINTS. COUNT RMS WEIGHT REMARK 3 RIGID-BOND RESTRAINTS (A**2): NULL ; NULL ; NULL REMARK 3 SPHERICITY; FREE ATOMS (A**2): NULL ; NULL ; NULL REMARK 3 SPHERICITY; BONDED ATOMS (A**2): NULL ; NULL ; NULL REMARK 3 REMARK 3 NCS RESTRAINTS STATISTICS REMARK 3 NUMBER OF DIFFERENT NCS GROUPS : NULL REMARK 3 REMARK 3 TLS DETAILS REMARK 3 NUMBER OF TLS GROUPS : 3 REMARK 3 REMARK 3 TLS GROUP : 1 REMARK 3 NUMBER OF COMPONENTS GROUP : 1 REMARK 3 COMPONENTS C SSSEQI TO C SSSEQI REMARK 3 RESIDUE RANGE : A 39 A 189 REMARK 3 ORIGIN FOR THE GROUP (A): 2.5006 14.8637 57.5834 REMARK 3 T TENSOR REMARK 3 T11: 0.1639 T22: 0.0488 REMARK 3 T33: 0.0415 T12: -0.0423 REMARK 3 T13: -0.0321 T23: -0.0093 REMARK 3 L TENSOR REMARK 3 L11: 1.4504 L22: 5.1696 REMARK 3 L33: 3.2553 L12: 0.1722 REMARK 3 L13: 0.7255 L23: -0.0322 REMARK 3 S TENSOR REMARK 3 S11: 0.0995 S12: -0.1308 S13: 0.0601 REMARK 3 S21: 0.3709 S22: -0.1246 S23: 0.2704 REMARK 3 S31: 0.2203 S32: -0.2641 S33: 0.0251 REMARK 3 REMARK 3 TLS GROUP : 2 REMARK 3 NUMBER OF COMPONENTS GROUP : 1 REMARK 3 COMPONENTS C SSSEQI TO C SSSEQI REMARK 3 RESIDUE RANGE : H 1 H 219 REMARK 3 ORIGIN FOR THE GROUP (A): -7.6258 12.1166 18.4022 REMARK 3 T TENSOR REMARK 3 T11: 0.0352 T22: 0.0326 REMARK 3 T33: 0.0113 T12: -0.0245 REMARK 3 T13: -0.0052 T23: 0.0131 REMARK 3 L TENSOR REMARK 3 L11: 1.5614 L22: 1.1648 REMARK 3 L33: 2.7818 L12: -0.0175 REMARK 3 L13: 1.1700 L23: -0.0447 REMARK 3 S TENSOR REMARK 3 S11: -0.0255 S12: 0.0345 S13: 0.0712 REMARK 3 S21: 0.1613 S22: -0.0574 S23: 0.0355 REMARK 3 S31: -0.0283 S32: 0.1623 S33: 0.0829 REMARK 3 REMARK 3 TLS GROUP : 3 REMARK 3 NUMBER OF COMPONENTS GROUP : 1 REMARK 3 COMPONENTS C SSSEQI TO C SSSEQI REMARK 3 RESIDUE RANGE : L 0 L 212 REMARK 3 ORIGIN FOR THE GROUP (A): -20.3410 -1.6659 21.6242 REMARK 3 T TENSOR REMARK 3 T11: 0.0875 T22: 0.0881 REMARK 3 T33: 0.0447 T12: -0.0771 REMARK 3 T13: -0.0234 T23: 0.0114 REMARK 3 L TENSOR REMARK 3 L11: 1.0526 L22: 0.4565 REMARK 3 L33: 2.0364 L12: 0.1440 REMARK 3 L13: 1.3809 L23: 0.3944 REMARK 3 S TENSOR REMARK 3 S11: 0.0949 S12: -0.0797 S13: 0.0062 REMARK 3 S21: 0.0845 S22: -0.1049 S23: 0.0962 REMARK 3 S31: 0.2216 S32: -0.1903 S33: 0.0099 REMARK 3 REMARK 3 BULK SOLVENT MODELLING. REMARK 3 METHOD USED : MASK REMARK 3 PARAMETERS FOR MASK CALCULATION REMARK 3 VDW PROBE RADIUS : 1.20 REMARK 3 ION PROBE RADIUS : 0.80 REMARK 3 SHRINKAGE RADIUS : 0.80 REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: HYDROGENS HAVE BEEN ADDED IN THE RIDING REMARK 3 POSITIONS REMARK 4 REMARK 4 8Z39 COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY PDBJ ON 15-APR-24. REMARK 100 THE DEPOSITION ID IS D_1300046917. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 18-DEC-18 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 : PHOTON FACTORY REMARK 200 BEAMLINE : BL-5A REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 1.0000 REMARK 200 MONOCHROMATOR : NULL REMARK 200 OPTICS : NULL REMARK 200 REMARK 200 DETECTOR TYPE : PIXEL REMARK 200 DETECTOR MANUFACTURER : DECTRIS PILATUS3 S 6M REMARK 200 INTENSITY-INTEGRATION SOFTWARE : XDS 20180409 REMARK 200 DATA SCALING SOFTWARE : AIMLESS 0.7.3 REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 70681 REMARK 200 RESOLUTION RANGE HIGH (A) : 1.800 REMARK 200 RESOLUTION RANGE LOW (A) : 48.650 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : NULL REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 99.9 REMARK 200 DATA REDUNDANCY : 5.100 REMARK 200 R MERGE (I) : 0.08800 REMARK 200 R SYM (I) : NULL REMARK 200 FOR THE DATA SET : 10.6000 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 1.80 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 1.84 REMARK 200 COMPLETENESS FOR SHELL (%) : 100.0 REMARK 200 DATA REDUNDANCY IN SHELL : 4.80 REMARK 200 R MERGE FOR SHELL (I) : 0.79000 REMARK 200 R SYM FOR SHELL (I) : NULL REMARK 200 FOR SHELL : 2.100 REMARK 200 REMARK 200 DIFFRACTION PROTOCOL: SINGLE WAVELENGTH REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: MOLECULAR REPLACEMENT REMARK 200 SOFTWARE USED: PHASER 2.8.2 REMARK 200 STARTING MODEL: NULL REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 54.52 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.70 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 200MM NACL, 20% PEG 3350, VAPOR REMARK 280 DIFFUSION, SITTING DROP, TEMPERATURE 293.15K REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: C 1 2 1 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 48.75950 REMARK 290 SMTRY2 3 0.000000 1.000000 0.000000 35.00250 REMARK 290 SMTRY3 3 0.000000 0.000000 1.000000 0.00000 REMARK 290 SMTRY1 4 -1.000000 0.000000 0.000000 48.75950 REMARK 290 SMTRY2 4 0.000000 1.000000 0.000000 35.00250 REMARK 290 SMTRY3 4 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: TRIMERIC REMARK 350 APPLY THE FOLLOWING TO CHAINS: A, 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 375 REMARK 375 SPECIAL POSITION REMARK 375 THE FOLLOWING ATOMS ARE FOUND TO BE WITHIN 0.15 ANGSTROMS REMARK 375 OF A SYMMETRY RELATED ATOM AND ARE ASSUMED TO BE ON SPECIAL REMARK 375 POSITIONS. REMARK 375 REMARK 375 ATOM RES CSSEQI REMARK 375 HOH H 480 LIES ON A SPECIAL POSITION. REMARK 375 HOH H 518 LIES ON A SPECIAL POSITION. 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 GLY A 29 REMARK 465 PHE A 30 REMARK 465 GLY A 31 REMARK 465 LYS A 32 REMARK 465 ARG A 33 REMARK 465 ARG A 34 REMARK 465 HIS A 35 REMARK 465 PRO A 36 REMARK 465 LYS A 37 REMARK 465 LYS A 38 REMARK 465 SER A 190 REMARK 465 VAL A 191 REMARK 465 ALA A 192 REMARK 465 ALA A 193 REMARK 465 LYS A 194 REMARK 465 SER A 195 REMARK 465 GLY A 196 REMARK 465 GLY A 197 REMARK 465 SER H 134 REMARK 465 LYS H 135 REMARK 465 SER H 136 REMARK 465 THR H 137 REMARK 465 SER H 138 REMARK 465 LYS H 220 REMARK 465 GLU H 221 REMARK 465 GLN H 222 REMARK 465 LYS H 223 REMARK 465 LEU H 224 REMARK 465 ILE H 225 REMARK 465 SER H 226 REMARK 465 GLU H 227 REMARK 465 GLU H 228 REMARK 465 ASP H 229 REMARK 465 LEU H 230 REMARK 465 ASN H 231 REMARK 465 SER H 232 REMARK 465 ALA H 233 REMARK 465 VAL H 234 REMARK 465 ASP H 235 REMARK 465 HIS H 236 REMARK 465 HIS H 237 REMARK 465 HIS H 238 REMARK 465 HIS H 239 REMARK 465 HIS H 240 REMARK 465 HIS H 241 REMARK 465 GLU L 213 REMARK 465 CYS L 214 REMARK 465 GLU L 215 REMARK 465 GLN L 216 REMARK 465 LYS L 217 REMARK 465 LEU L 218 REMARK 465 ILE L 219 REMARK 465 SER L 220 REMARK 465 GLU L 221 REMARK 465 GLU L 222 REMARK 465 ASP L 223 REMARK 465 LEU L 224 REMARK 465 ASN L 225 REMARK 465 SER L 226 REMARK 465 ALA L 227 REMARK 465 VAL L 228 REMARK 465 ASP L 229 REMARK 465 HIS L 230 REMARK 465 HIS L 231 REMARK 465 HIS L 232 REMARK 465 HIS L 233 REMARK 465 HIS L 234 REMARK 465 HIS L 235 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 A 509 O HOH A 608 2.12 REMARK 500 REMARK 500 REMARK: NULL REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: COVALENT BOND LENGTHS 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,2(A3,1X,A1,I4,A1,1X,A4,3X),1X,F6.3) 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 RES CSSEQI ATM2 DEVIATION REMARK 500 ASN A 50 CG ASN A 50 OD1 0.137 REMARK 500 GLU H 101 CD GLU H 101 OE1 0.146 REMARK 500 GLU H 154 CD GLU H 154 OE2 0.104 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 A 50 30.16 -65.67 REMARK 500 ALA A 58 -113.53 -106.67 REMARK 500 SER H 44 -133.58 54.32 REMARK 500 ASP H 150 58.63 75.67 REMARK 500 SER L 30 -124.40 50.46 REMARK 500 ALA L 51 -33.84 67.80 REMARK 500 REMARK 500 REMARK: NULL REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: NON-CIS, NON-TRANS REMARK 500 REMARK 500 THE FOLLOWING PEPTIDE BONDS DEVIATE SIGNIFICANTLY FROM BOTH REMARK 500 CIS AND TRANS CONFORMATION. CIS BONDS, IF ANY, ARE LISTED REMARK 500 ON CISPEP RECORDS. TRANS IS DEFINED AS 180 +/- 30 AND REMARK 500 CIS IS DEFINED AS 0 +/- 30 DEGREES. REMARK 500 MODEL OMEGA REMARK 500 PHE H 105 PHE H 106 -148.40 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 H 536 DISTANCE = 6.09 ANGSTROMS REMARK 525 HOH L 555 DISTANCE = 6.05 ANGSTROMS REMARK 620 REMARK 620 METAL COORDINATION REMARK 620 (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN IDENTIFIER; REMARK 620 SSEQ=SEQUENCE NUMBER; I=INSERTION CODE): REMARK 620 REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL REMARK 620 CA A 401 CA REMARK 620 N RES CSSEQI ATOM REMARK 620 1 GLU A 89 OE1 REMARK 620 2 GLU A 89 OE2 54.5 REMARK 620 3 GLU A 90 OE1 73.4 80.6 REMARK 620 4 GLU A 90 OE2 111.3 78.5 49.8 REMARK 620 5 ASP A 95 OD1 139.2 160.4 90.4 82.3 REMARK 620 6 ASP A 95 OD2 88.0 141.7 81.3 113.5 52.1 REMARK 620 7 THR A 125 O 94.8 107.3 158.6 150.2 87.3 80.5 REMARK 620 8 GLU A 126 OE2 134.4 82.4 117.5 68.0 86.4 135.9 83.6 REMARK 620 N 1 2 3 4 5 6 7 REMARK 620 REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL REMARK 620 CA A 402 CA REMARK 620 N RES CSSEQI ATOM REMARK 620 1 GLU A 90 OE2 REMARK 620 2 GLU A 126 OE1 113.6 REMARK 620 3 GLU A 126 OE2 65.0 48.9 REMARK 620 4 ASP A 129 OD1 157.2 87.8 136.7 REMARK 620 5 ASP A 131 OD2 90.1 141.1 136.2 76.5 REMARK 620 6 HOH A 512 O 75.1 86.9 79.6 99.3 130.3 REMARK 620 7 HOH A 573 O 90.5 74.2 69.9 103.6 75.3 149.5 REMARK 620 8 HOH A 585 O 89.2 140.0 142.3 68.5 65.7 66.9 140.9 REMARK 620 N 1 2 3 4 5 6 7 REMARK 620 REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL REMARK 620 ZN A 403 ZN REMARK 620 N RES CSSEQI ATOM REMARK 620 1 HIS A 140 NE2 REMARK 620 2 ASP A 147 OD1 116.0 REMARK 620 3 HIS A 182 ND1 107.4 107.1 REMARK 620 4 SO4 A 405 O1 83.0 132.7 107.1 REMARK 620 5 HOH A 506 O 115.7 105.3 104.5 34.2 REMARK 620 N 1 2 3 4 DBREF 8Z39 A 29 197 UNP Q15465 SHH_HUMAN 29 197 DBREF 8Z39 H 1 241 PDB 8Z39 8Z39 1 241 DBREF 8Z39 L 0 235 PDB 8Z39 8Z39 0 235 SEQRES 1 A 169 GLY PHE GLY LYS ARG ARG HIS PRO LYS LYS LEU THR PRO SEQRES 2 A 169 LEU ALA TYR LYS GLN PHE ILE PRO ASN VAL ALA GLU LYS SEQRES 3 A 169 THR LEU GLY ALA SER GLY ARG TYR GLU GLY LYS ILE SER SEQRES 4 A 169 ARG ASN SER GLU ARG PHE LYS GLU LEU THR PRO ASN TYR SEQRES 5 A 169 ASN PRO ASP ILE ILE PHE LYS ASP GLU GLU ASN THR GLY SEQRES 6 A 169 ALA ASP ARG LEU MET THR GLN ARG CYS LYS ASP LYS LEU SEQRES 7 A 169 ASN ALA LEU ALA ILE SER VAL MET ASN GLN TRP PRO GLY SEQRES 8 A 169 VAL LYS LEU ARG VAL THR GLU GLY TRP ASP GLU ASP GLY SEQRES 9 A 169 HIS HIS SER GLU GLU SER LEU HIS TYR GLU GLY ARG ALA SEQRES 10 A 169 VAL ASP ILE THR THR SER ASP ARG ASP ARG SER LYS TYR SEQRES 11 A 169 GLY MET LEU ALA ARG LEU ALA VAL GLU ALA GLY PHE ASP SEQRES 12 A 169 TRP VAL TYR TYR GLU SER LYS ALA HIS ILE HIS CYS SER SEQRES 13 A 169 VAL LYS ALA GLU ASN SER VAL ALA ALA LYS SER GLY GLY SEQRES 1 H 241 GLN VAL GLN LEU GLN GLN SER GLY PRO GLU LEU VAL ARG SEQRES 2 H 241 PRO GLY VAL SER VAL LYS ILE SER CYS LYS GLY SER GLY SEQRES 3 H 241 TYR THR PHE ILE ASP GLU ALA LEU HIS TRP VAL LYS GLN SEQRES 4 H 241 SER HIS ALA GLU SER LEU GLU TRP ILE GLY VAL ILE ARG SEQRES 5 H 241 PRO TYR SER GLY GLU THR ASN TYR ASN GLN LYS PHE LYS SEQRES 6 H 241 ASP LYS ALA THR MET THR VAL ASP ILE SER SER SER THR SEQRES 7 H 241 ALA TYR LEU GLU LEU ALA ARG LEU THR SER GLU ASP SER SEQRES 8 H 241 ALA ILE TYR TYR CYS ALA ARG ASP TRP GLU ARG GLY ASP SEQRES 9 H 241 PHE PHE ASP TYR TRP GLY GLN GLY THR LEU VAL THR VAL SEQRES 10 H 241 SER SER ALA SER THR LYS GLY PRO SER VAL PHE PRO LEU SEQRES 11 H 241 ALA PRO SER SER LYS SER THR SER GLY GLY THR ALA ALA SEQRES 12 H 241 LEU GLY CYS LEU VAL LYS ASP TYR PHE PRO GLU PRO VAL SEQRES 13 H 241 THR VAL SER TRP ASN SER GLY ALA LEU THR SER GLY VAL SEQRES 14 H 241 HIS THR PHE PRO ALA VAL LEU GLN SER SER GLY LEU TYR SEQRES 15 H 241 SER LEU SER SER VAL VAL THR VAL PRO SER SER SER LEU SEQRES 16 H 241 GLY THR GLN THR TYR ILE CYS ASN VAL ASN HIS LYS PRO SEQRES 17 H 241 SER ASN THR LYS VAL ASP LYS LYS VAL GLU PRO LYS GLU SEQRES 18 H 241 GLN LYS LEU ILE SER GLU GLU ASP LEU ASN SER ALA VAL SEQRES 19 H 241 ASP HIS HIS HIS HIS HIS HIS SEQRES 1 L 236 GLY ASP ILE VAL MET THR GLN THR PRO LYS PHE LEU LEU SEQRES 2 L 236 VAL SER ALA GLY ASP LYS VAL THR ILE THR CYS LYS ALA SEQRES 3 L 236 SER GLN SER VAL SER ASN ASP LEU THR TRP TYR GLN GLN SEQRES 4 L 236 LYS PRO GLY GLN SER PRO LYS LEU LEU ILE TYR TYR ALA SEQRES 5 L 236 SER ASN ARG TYR ALA GLY VAL PRO ASP ARG PHE THR GLY SEQRES 6 L 236 SER GLY TYR GLY THR ASP PHE THR PHE THR ILE SER THR SEQRES 7 L 236 VAL GLN ALA GLU ASP LEU ALA VAL TYR PHE CYS GLN GLN SEQRES 8 L 236 ASP TYR GLY SER PRO PRO THR PHE GLY GLY GLY THR LYS SEQRES 9 L 236 VAL GLU ILE LYS ARG THR VAL ALA ALA PRO SER VAL PHE SEQRES 10 L 236 ILE PHE PRO PRO SER ASP GLU GLN LEU LYS SER GLY THR SEQRES 11 L 236 ALA SER VAL VAL CYS LEU LEU ASN ASN PHE TYR PRO ARG SEQRES 12 L 236 GLU ALA LYS VAL GLN TRP LYS VAL ASP ASN ALA LEU GLN SEQRES 13 L 236 SER GLY ASN SER GLN GLU SER VAL THR GLU GLN ASP SER SEQRES 14 L 236 LYS ASP SER THR TYR SER LEU SER SER THR LEU THR LEU SEQRES 15 L 236 SER LYS ALA ASP TYR GLU LYS HIS LYS VAL TYR ALA CYS SEQRES 16 L 236 GLU VAL THR HIS GLN GLY LEU SER SER PRO VAL THR LYS SEQRES 17 L 236 SER PHE ASN ARG GLY GLU CYS GLU GLN LYS LEU ILE SER SEQRES 18 L 236 GLU GLU ASP LEU ASN SER ALA VAL ASP HIS HIS HIS HIS SEQRES 19 L 236 HIS HIS HET CA A 401 1 HET CA A 402 1 HET ZN A 403 1 HET CL A 404 1 HET SO4 A 405 5 HETNAM CA CALCIUM ION HETNAM ZN ZINC ION HETNAM CL CHLORIDE ION HETNAM SO4 SULFATE ION FORMUL 4 CA 2(CA 2+) FORMUL 6 ZN ZN 2+ FORMUL 7 CL CL 1- FORMUL 8 SO4 O4 S 2- FORMUL 9 HOH *625(H2 O) HELIX 1 AA1 GLU A 71 LEU A 76 5 6 HELIX 2 AA2 GLY A 93 ASP A 95 5 3 HELIX 3 AA3 THR A 99 TRP A 117 1 19 HELIX 4 AA4 SER A 138 GLY A 143 5 6 HELIX 5 AA5 ASP A 154 SER A 156 5 3 HELIX 6 AA6 LYS A 157 ALA A 168 1 12 HELIX 7 AA7 THR H 28 GLU H 32 5 5 HELIX 8 AA8 GLN H 62 LYS H 65 5 4 HELIX 9 AA9 ILE H 74 SER H 76 5 3 HELIX 10 AB1 THR H 87 SER H 91 5 5 HELIX 11 AB2 SER H 162 ALA H 164 5 3 HELIX 12 AB3 SER H 193 LEU H 195 5 3 HELIX 13 AB4 LYS H 207 ASN H 210 5 4 HELIX 14 AB5 GLN L 79 LEU L 83 5 5 HELIX 15 AB6 SER L 121 SER L 127 1 7 HELIX 16 AB7 LYS L 183 GLU L 187 1 5 SHEET 1 AA1 6 PHE A 47 ILE A 48 0 SHEET 2 AA1 6 TRP A 172 SER A 177 -1 O VAL A 173 N ILE A 48 SHEET 3 AA1 6 HIS A 180 SER A 184 -1 O HIS A 182 N TYR A 174 SHEET 4 AA1 6 ALA A 145 THR A 150 -1 N VAL A 146 O CYS A 183 SHEET 5 AA1 6 LEU A 122 GLU A 126 -1 N GLU A 126 O ASP A 147 SHEET 6 AA1 6 ILE A 84 PHE A 86 1 N ILE A 85 O VAL A 124 SHEET 1 AA2 2 THR A 77 PRO A 78 0 SHEET 2 AA2 2 LEU A 97 MET A 98 -1 O MET A 98 N THR A 77 SHEET 1 AA3 4 GLN H 3 GLN H 6 0 SHEET 2 AA3 4 VAL H 18 SER H 25 -1 O LYS H 23 N GLN H 5 SHEET 3 AA3 4 THR H 78 LEU H 83 -1 O ALA H 79 N CYS H 22 SHEET 4 AA3 4 ALA H 68 ASP H 73 -1 N ASP H 73 O THR H 78 SHEET 1 AA4 6 GLU H 10 VAL H 12 0 SHEET 2 AA4 6 THR H 113 VAL H 117 1 O THR H 116 N GLU H 10 SHEET 3 AA4 6 ALA H 92 ARG H 98 -1 N ALA H 92 O VAL H 115 SHEET 4 AA4 6 LEU H 34 SER H 40 -1 N HIS H 35 O ALA H 97 SHEET 5 AA4 6 SER H 44 ILE H 51 -1 O GLU H 46 N LYS H 38 SHEET 6 AA4 6 THR H 58 TYR H 60 -1 O ASN H 59 N VAL H 50 SHEET 1 AA5 4 GLU H 10 VAL H 12 0 SHEET 2 AA5 4 THR H 113 VAL H 117 1 O THR H 116 N GLU H 10 SHEET 3 AA5 4 ALA H 92 ARG H 98 -1 N ALA H 92 O VAL H 115 SHEET 4 AA5 4 TYR H 108 TRP H 109 -1 O TYR H 108 N ARG H 98 SHEET 1 AA6 4 SER H 126 LEU H 130 0 SHEET 2 AA6 4 THR H 141 TYR H 151 -1 O LYS H 149 N SER H 126 SHEET 3 AA6 4 TYR H 182 PRO H 191 -1 O LEU H 184 N VAL H 148 SHEET 4 AA6 4 VAL H 169 THR H 171 -1 N HIS H 170 O VAL H 187 SHEET 1 AA7 4 SER H 126 LEU H 130 0 SHEET 2 AA7 4 THR H 141 TYR H 151 -1 O LYS H 149 N SER H 126 SHEET 3 AA7 4 TYR H 182 PRO H 191 -1 O LEU H 184 N VAL H 148 SHEET 4 AA7 4 VAL H 175 LEU H 176 -1 N VAL H 175 O SER H 183 SHEET 1 AA8 3 THR H 157 TRP H 160 0 SHEET 2 AA8 3 ILE H 201 HIS H 206 -1 O ASN H 203 N SER H 159 SHEET 3 AA8 3 THR H 211 LYS H 216 -1 O THR H 211 N HIS H 206 SHEET 1 AA9 4 MET L 4 THR L 7 0 SHEET 2 AA9 4 VAL L 19 ALA L 25 -1 O THR L 22 N THR L 7 SHEET 3 AA9 4 ASP L 70 ILE L 75 -1 O PHE L 71 N CYS L 23 SHEET 4 AA9 4 PHE L 62 GLY L 66 -1 N SER L 65 O THR L 72 SHEET 1 AB1 6 PHE L 10 SER L 14 0 SHEET 2 AB1 6 THR L 102 LYS L 107 1 O GLU L 105 N LEU L 11 SHEET 3 AB1 6 ALA L 84 GLN L 90 -1 N ALA L 84 O VAL L 104 SHEET 4 AB1 6 LEU L 33 GLN L 38 -1 N TYR L 36 O PHE L 87 SHEET 5 AB1 6 LYS L 45 TYR L 49 -1 O LEU L 47 N TRP L 35 SHEET 6 AB1 6 ASN L 53 ARG L 54 -1 O ASN L 53 N TYR L 49 SHEET 1 AB2 4 PHE L 10 SER L 14 0 SHEET 2 AB2 4 THR L 102 LYS L 107 1 O GLU L 105 N LEU L 11 SHEET 3 AB2 4 ALA L 84 GLN L 90 -1 N ALA L 84 O VAL L 104 SHEET 4 AB2 4 THR L 97 PHE L 98 -1 O THR L 97 N GLN L 90 SHEET 1 AB3 4 SER L 114 PHE L 118 0 SHEET 2 AB3 4 THR L 129 PHE L 139 -1 O VAL L 133 N PHE L 118 SHEET 3 AB3 4 TYR L 173 SER L 182 -1 O TYR L 173 N PHE L 139 SHEET 4 AB3 4 SER L 159 VAL L 163 -1 N GLN L 160 O THR L 178 SHEET 1 AB4 4 ALA L 153 LEU L 154 0 SHEET 2 AB4 4 LYS L 145 VAL L 150 -1 N VAL L 150 O ALA L 153 SHEET 3 AB4 4 VAL L 191 THR L 197 -1 O GLU L 195 N GLN L 147 SHEET 4 AB4 4 VAL L 205 ASN L 210 -1 O VAL L 205 N VAL L 196 SSBOND 1 CYS H 22 CYS H 96 1555 1555 2.09 SSBOND 2 CYS H 146 CYS H 202 1555 1555 2.03 SSBOND 3 CYS L 23 CYS L 88 1555 1555 2.17 SSBOND 4 CYS L 134 CYS L 194 1555 1555 1.99 LINK OE1 GLU A 89 CA CA A 401 1555 1555 2.40 LINK OE2 GLU A 89 CA CA A 401 1555 1555 2.38 LINK OE1 GLU A 90 CA CA A 401 1555 1555 2.63 LINK OE2 GLU A 90 CA CA A 401 1555 1555 2.52 LINK OE2 GLU A 90 CA CA A 402 1555 1555 2.34 LINK OD1 ASP A 95 CA CA A 401 1555 1555 2.55 LINK OD2 ASP A 95 CA CA A 401 1555 1555 2.47 LINK O THR A 125 CA CA A 401 1555 1555 2.28 LINK OE2 GLU A 126 CA CA A 401 1555 1555 2.30 LINK OE1 GLU A 126 CA CA A 402 1555 1555 2.49 LINK OE2 GLU A 126 CA CA A 402 1555 1555 2.66 LINK OD1 ASP A 129 CA CA A 402 1555 1555 2.33 LINK OD2 ASP A 131 CA CA A 402 1555 1555 2.53 LINK NE2 HIS A 140 ZN ZN A 403 1555 1555 1.93 LINK OD1 ASP A 147 ZN ZN A 403 1555 1555 1.94 LINK ND1 HIS A 182 ZN ZN A 403 1555 1555 2.03 LINK CA CA A 402 O HOH A 512 1555 1555 2.48 LINK CA CA A 402 O HOH A 573 1555 1555 2.42 LINK CA CA A 402 O HOH A 585 1555 1555 3.19 LINK ZN ZN A 403 O1 BSO4 A 405 1555 1555 2.31 LINK ZN ZN A 403 O AHOH A 506 1555 1555 2.10 CISPEP 1 ILE A 48 PRO A 49 0 1.78 CISPEP 2 PHE H 152 PRO H 153 0 -5.89 CISPEP 3 GLU H 154 PRO H 155 0 -8.01 CISPEP 4 THR L 7 PRO L 8 0 -6.54 CISPEP 5 SER L 94 PRO L 95 0 0.53 CISPEP 6 TYR L 140 PRO L 141 0 -3.86 CRYST1 97.519 70.005 119.974 90.00 109.24 90.00 C 1 2 1 4 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 0.010254 0.000000 0.003580 0.00000 SCALE2 0.000000 0.014285 0.000000 0.00000 SCALE3 0.000000 0.000000 0.008828 0.00000