HEADER IMMUNE SYSTEM 12-APR-24 9EZH TITLE STRUCTURE OF SINGLE-DOMAIN ANTIBODY VHH_H4 IN COMPLEX WITH HUMAN VSIG4 COMPND MOL_ID: 1; COMPND 2 MOLECULE: V-SET AND IMMUNOGLOBULIN DOMAIN-CONTAINING PROTEIN 4; COMPND 3 CHAIN: A; COMPND 4 SYNONYM: PROTEIN Z39IG; COMPND 5 ENGINEERED: YES; COMPND 6 MOL_ID: 2; COMPND 7 MOLECULE: VHH_H4; COMPND 8 CHAIN: B; COMPND 9 ENGINEERED: YES; COMPND 10 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: VSIG4, CRIG, Z39IG, UNQ317/PRO362; SOURCE 6 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 7 EXPRESSION_SYSTEM_TAXID: 562; SOURCE 8 MOL_ID: 2; SOURCE 9 ORGANISM_SCIENTIFIC: CAMELIDAE; SOURCE 10 ORGANISM_TAXID: 9835; SOURCE 11 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 12 EXPRESSION_SYSTEM_TAXID: 562 KEYWDS NANOBODY, MUTATIONS, CDR2, CDR3, HVSIG4, B7 FAMILY, IMMUNE REGULATORY KEYWDS 2 PROTEINS, IMMUNE SYSTEM EXPDTA X-RAY DIFFRACTION AUTHOR D.VIZARRAGA,D.CIANFERONI,J.DELGADO,R.VAN DER KANT,T.GARCIA, AUTHOR 2 K.MARAGKOU,Z.ZHANG,M.DEWILDE,J.SCHYMKOWITZ,F.ROUSSEAU,L.SERRANO REVDAT 1 29-OCT-25 9EZH 0 JRNL AUTH R.VAN DER KANT,Z.ZHANG,I.MARKOVIC,D.VIZARRAGA,T.GARCIA, JRNL AUTH 2 K.MARAGKOU,J.DELGADO,D.CIANFERONI,G.ORLANDO,C.CAROLIS, JRNL AUTH 3 S.SAVVIDES,A.N.VOLKOV,M.DEWILDE,J.SCHYMKOWITZ,L.SERRANO, JRNL AUTH 4 F.ROUSSEAU JRNL TITL IN SILICO DESIGN OF STABLE SINGLE-DOMAIN ANTIBODIES AGAINST JRNL TITL 2 MULTIPLE TARGETS. JRNL REF TO BE PUBLISHED JRNL REFN REMARK 2 REMARK 2 RESOLUTION. 1.94 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : PHENIX (1.20_4459: ???) 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 : ML REMARK 3 REMARK 3 DATA USED IN REFINEMENT. REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 1.94 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 48.93 REMARK 3 MIN(FOBS/SIGMA_FOBS) : 1.350 REMARK 3 COMPLETENESS FOR RANGE (%) : 99.9 REMARK 3 NUMBER OF REFLECTIONS : 18055 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 R VALUE (WORKING + TEST SET) : 0.196 REMARK 3 R VALUE (WORKING SET) : 0.194 REMARK 3 FREE R VALUE : 0.239 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 4.600 REMARK 3 FREE R VALUE TEST SET COUNT : 830 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 48.9300 - 3.5200 1.00 3049 163 0.1777 0.2150 REMARK 3 2 3.5200 - 2.7900 1.00 2870 141 0.1896 0.2407 REMARK 3 3 2.7900 - 2.4400 1.00 2859 141 0.2139 0.2834 REMARK 3 4 2.4400 - 2.2200 1.00 2827 131 0.2100 0.2546 REMARK 3 5 2.2200 - 2.0600 1.00 2806 130 0.2244 0.2586 REMARK 3 6 2.0600 - 1.9400 1.00 2814 124 0.2408 0.3093 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) : 0.200 REMARK 3 PHASE ERROR (DEGREES, MAXIMUM-LIKELIHOOD BASED) : 26.750 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : NULL REMARK 3 MEAN B VALUE (OVERALL, A**2) : NULL REMARK 3 OVERALL ANISOTROPIC B VALUE. REMARK 3 B11 (A**2) : NULL REMARK 3 B22 (A**2) : NULL REMARK 3 B33 (A**2) : NULL REMARK 3 B12 (A**2) : NULL REMARK 3 B13 (A**2) : NULL REMARK 3 B23 (A**2) : NULL REMARK 3 REMARK 3 TWINNING INFORMATION. REMARK 3 FRACTION: NULL REMARK 3 OPERATOR: NULL REMARK 3 REMARK 3 DEVIATIONS FROM IDEAL VALUES. REMARK 3 RMSD COUNT REMARK 3 BOND : 0.008 NULL REMARK 3 ANGLE : 1.057 NULL REMARK 3 CHIRALITY : 0.066 281 REMARK 3 PLANARITY : 0.009 360 REMARK 3 DIHEDRAL : 6.710 280 REMARK 3 REMARK 3 TLS DETAILS REMARK 3 NUMBER OF TLS GROUPS : 1 REMARK 3 TLS GROUP : 1 REMARK 3 SELECTION: ALL REMARK 3 ORIGIN FOR THE GROUP (A): -2.1082 -9.2444 22.6379 REMARK 3 T TENSOR REMARK 3 T11: 0.3900 T22: 0.3078 REMARK 3 T33: 0.2710 T12: 0.0706 REMARK 3 T13: -0.0310 T23: 0.0230 REMARK 3 L TENSOR REMARK 3 L11: 1.4033 L22: 3.1704 REMARK 3 L33: 3.7641 L12: -0.3097 REMARK 3 L13: -0.4823 L23: 1.1322 REMARK 3 S TENSOR REMARK 3 S11: 0.1366 S12: 0.3822 S13: 0.1381 REMARK 3 S21: -0.7113 S22: -0.2051 S23: 0.1297 REMARK 3 S31: -0.7884 S32: -0.2395 S33: 0.0316 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 9EZH COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY PDBE ON 12-APR-24. REMARK 100 THE DEPOSITION ID IS D_1292136122. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 15-JUL-23 REMARK 200 TEMPERATURE (KELVIN) : 80.0 REMARK 200 PH : NULL REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : Y REMARK 200 RADIATION SOURCE : ALBA REMARK 200 BEAMLINE : XALOC REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 0.97926 REMARK 200 MONOCHROMATOR : NULL REMARK 200 OPTICS : NULL REMARK 200 REMARK 200 DETECTOR TYPE : PIXEL REMARK 200 DETECTOR MANUFACTURER : DECTRIS PILATUS 6M REMARK 200 INTENSITY-INTEGRATION SOFTWARE : XDS REMARK 200 DATA SCALING SOFTWARE : AIMLESS REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 18076 REMARK 200 RESOLUTION RANGE HIGH (A) : 1.938 REMARK 200 RESOLUTION RANGE LOW (A) : 87.619 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : NULL REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 100.0 REMARK 200 DATA REDUNDANCY : 12.60 REMARK 200 R MERGE (I) : 0.09600 REMARK 200 R SYM (I) : NULL REMARK 200 FOR THE DATA SET : 16.0000 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 1.94 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 1.97 REMARK 200 COMPLETENESS FOR SHELL (%) : NULL REMARK 200 DATA REDUNDANCY IN SHELL : NULL REMARK 200 R MERGE FOR SHELL (I) : 1.09400 REMARK 200 R SYM FOR SHELL (I) : NULL REMARK 200 FOR SHELL : 2.500 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 (%): 42.64 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.14 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 0,1M BIS-TRIS PH 6.0 AND 24% PEG3350, REMARK 280 VAPOR DIFFUSION, HANGING DROP, TEMPERATURE 293K REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 21 21 21 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 -X+1/2,-Y,Z+1/2 REMARK 290 3555 -X,Y+1/2,-Z+1/2 REMARK 290 4555 X+1/2,-Y+1/2,-Z REMARK 290 REMARK 290 WHERE NNN -> OPERATOR NUMBER REMARK 290 MMM -> TRANSLATION VECTOR REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY TRANSFORMATIONS REMARK 290 THE FOLLOWING TRANSFORMATIONS OPERATE ON THE ATOM/HETATM REMARK 290 RECORDS IN THIS ENTRY TO PRODUCE CRYSTALLOGRAPHICALLY REMARK 290 RELATED MOLECULES. REMARK 290 SMTRY1 1 1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 1 0.000000 1.000000 0.000000 0.00000 REMARK 290 SMTRY3 1 0.000000 0.000000 1.000000 0.00000 REMARK 290 SMTRY1 2 -1.000000 0.000000 0.000000 12.88200 REMARK 290 SMTRY2 2 0.000000 -1.000000 0.000000 0.00000 REMARK 290 SMTRY3 2 0.000000 0.000000 1.000000 87.61900 REMARK 290 SMTRY1 3 -1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 3 0.000000 1.000000 0.000000 25.48050 REMARK 290 SMTRY3 3 0.000000 0.000000 -1.000000 87.61900 REMARK 290 SMTRY1 4 1.000000 0.000000 0.000000 12.88200 REMARK 290 SMTRY2 4 0.000000 -1.000000 0.000000 25.48050 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: DIMERIC REMARK 350 SOFTWARE DETERMINED QUATERNARY STRUCTURE: DIMERIC REMARK 350 SOFTWARE USED: PISA REMARK 350 TOTAL BURIED SURFACE AREA: 1810 ANGSTROM**2 REMARK 350 SURFACE AREA OF THE COMPLEX: 11360 ANGSTROM**2 REMARK 350 CHANGE IN SOLVENT FREE ENERGY: 0.0 KCAL/MOL 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 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 352 O HOH A 383 2.17 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 ASP A 52 -155.58 -132.67 REMARK 500 HIS A 71 -10.66 -147.86 REMARK 500 SER A 90 169.54 175.83 REMARK 500 THR B 90 103.89 -53.46 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 393 DISTANCE = 6.55 ANGSTROMS REMARK 525 HOH B 223 DISTANCE = 7.65 ANGSTROMS REMARK 525 HOH B 224 DISTANCE = 7.98 ANGSTROMS DBREF 9EZH A 1 118 UNP Q9Y279 VSIG4_HUMAN 20 137 DBREF 9EZH B 1 121 PDB 9EZH 9EZH 1 121 SEQRES 1 A 118 ARG PRO ILE LEU GLU VAL PRO GLU SER VAL THR GLY PRO SEQRES 2 A 118 TRP LYS GLY ASP VAL ASN LEU PRO CYS THR TYR ASP PRO SEQRES 3 A 118 LEU GLN GLY TYR THR GLN VAL LEU VAL LYS TRP LEU VAL SEQRES 4 A 118 GLN ARG GLY SER ASP PRO VAL THR ILE PHE LEU ARG ASP SEQRES 5 A 118 SER SER GLY ASP HIS ILE GLN GLN ALA LYS TYR GLN GLY SEQRES 6 A 118 ARG LEU HIS VAL SER HIS LYS VAL PRO GLY ASP VAL SER SEQRES 7 A 118 LEU GLN LEU SER THR LEU GLU MET ASP ASP ARG SER HIS SEQRES 8 A 118 TYR THR CYS GLU VAL THR TRP GLN THR PRO ASP GLY ASN SEQRES 9 A 118 GLN VAL VAL ARG ASP LYS ILE THR GLU LEU ARG VAL GLN SEQRES 10 A 118 LYS SEQRES 1 B 121 VAL GLN LEU VAL GLU SER GLY GLY GLY LEU VAL GLN ALA SEQRES 2 B 121 GLY GLY SER LEU ARG LEU SER CYS ALA ALA SER GLY ARG SEQRES 3 B 121 THR PHE SER SER TYR GLY MET GLY TRP PHE ARG GLN ALA SEQRES 4 B 121 PRO GLY LYS GLU ARG GLU PHE VAL ALA ALA ILE GLY PRO SEQRES 5 B 121 PHE GLY GLY HIS THR TYR TYR ALA ASP SER VAL LYS GLY SEQRES 6 B 121 ARG PHE THR ILE SER ARG ASP ASN ALA LYS ASN THR VAL SEQRES 7 B 121 TYR LEU GLN MET ASN SER LEU LYS PRO GLU ASP THR ALA SEQRES 8 B 121 VAL TYR TYR CYS ALA ALA GLY ARG TRP ASN ARG TYR GLY SEQRES 9 B 121 PHE TYR ASP GLN ASP ASN TYR ASP TYR TRP GLY GLN GLY SEQRES 10 B 121 THR GLN VAL THR HET GOL A 201 6 HETNAM GOL GLYCEROL HETSYN GOL GLYCERIN; PROPANE-1,2,3-TRIOL FORMUL 3 GOL C3 H8 O3 FORMUL 4 HOH *117(H2 O) HELIX 1 AA1 GLU A 85 ARG A 89 5 5 HELIX 2 AA2 THR B 27 TYR B 31 5 5 HELIX 3 AA3 LYS B 86 THR B 90 5 5 HELIX 4 AA4 ASP B 107 TYR B 111 5 5 SHEET 1 AA1 6 SER A 9 PRO A 13 0 SHEET 2 AA1 6 GLN A 105 GLN A 117 1 O ARG A 115 N VAL A 10 SHEET 3 AA1 6 SER A 90 GLN A 99 -1 N SER A 90 O LEU A 114 SHEET 4 AA1 6 THR A 31 ARG A 41 -1 N GLN A 40 O HIS A 91 SHEET 5 AA1 6 ASP A 44 ASP A 52 -1 O VAL A 46 N VAL A 39 SHEET 6 AA1 6 GLY A 55 ILE A 58 -1 O HIS A 57 N LEU A 50 SHEET 1 AA2 3 VAL A 18 LEU A 20 0 SHEET 2 AA2 3 LEU A 79 LEU A 81 -1 O LEU A 79 N LEU A 20 SHEET 3 AA2 3 LEU A 67 VAL A 69 -1 N HIS A 68 O GLN A 80 SHEET 1 AA3 4 GLN B 2 SER B 6 0 SHEET 2 AA3 4 SER B 16 SER B 24 -1 O SER B 24 N GLN B 2 SHEET 3 AA3 4 THR B 77 ASN B 83 -1 O LEU B 80 N LEU B 19 SHEET 4 AA3 4 PHE B 67 ASP B 72 -1 N THR B 68 O GLN B 81 SHEET 1 AA4 5 THR B 57 TYR B 59 0 SHEET 2 AA4 5 GLU B 45 ILE B 50 -1 N ALA B 49 O TYR B 58 SHEET 3 AA4 5 GLY B 32 GLN B 38 -1 N ARG B 37 O GLU B 45 SHEET 4 AA4 5 ALA B 91 GLY B 98 -1 O TYR B 94 N PHE B 36 SHEET 5 AA4 5 TYR B 113 TRP B 114 -1 O TYR B 113 N ALA B 97 SHEET 1 AA5 5 THR B 57 TYR B 59 0 SHEET 2 AA5 5 GLU B 45 ILE B 50 -1 N ALA B 49 O TYR B 58 SHEET 3 AA5 5 GLY B 32 GLN B 38 -1 N ARG B 37 O GLU B 45 SHEET 4 AA5 5 ALA B 91 GLY B 98 -1 O TYR B 94 N PHE B 36 SHEET 5 AA5 5 THR B 118 VAL B 120 -1 O THR B 118 N TYR B 93 SSBOND 1 CYS A 22 CYS A 94 1555 1555 2.03 SSBOND 2 CYS B 21 CYS B 95 1555 1555 2.04 CISPEP 1 ARG A 1 PRO A 2 0 -7.69 CRYST1 25.764 50.961 175.238 90.00 90.00 90.00 P 21 21 21 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 0.038814 0.000000 0.000000 0.00000 SCALE2 0.000000 0.019623 0.000000 0.00000 SCALE3 0.000000 0.000000 0.005707 0.00000