HEADER VIRAL PROTEIN/IMMUNE SYSTEM 27-JUN-24 9CFH TITLE CRYO-EM REFINEMENT OF ANTIBODY 19-77 R71V IN COMPLEX WITH SARS-COV-2 TITLE 2 JD.1.1 RBD COMPND MOL_ID: 1; COMPND 2 MOLECULE: SPIKE PROTEIN S1; COMPND 3 CHAIN: A; COMPND 4 ENGINEERED: YES; COMPND 5 MOL_ID: 2; COMPND 6 MOLECULE: HEAVY CHAIN; COMPND 7 CHAIN: H; COMPND 8 ENGINEERED: YES; COMPND 9 MOL_ID: 3; COMPND 10 MOLECULE: LIGHT CHAIN; COMPND 11 CHAIN: L; COMPND 12 ENGINEERED: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: SEVERE ACUTE RESPIRATORY SYNDROME CORONAVIRUS SOURCE 3 2; SOURCE 4 ORGANISM_TAXID: 2697049; SOURCE 5 GENE: S, 2; SOURCE 6 EXPRESSION_SYSTEM: HOMO SAPIENS; SOURCE 7 EXPRESSION_SYSTEM_TAXID: 9606; SOURCE 8 MOL_ID: 2; SOURCE 9 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 10 ORGANISM_TAXID: 9606; SOURCE 11 EXPRESSION_SYSTEM: HOMO SAPIENS; SOURCE 12 EXPRESSION_SYSTEM_TAXID: 9606; SOURCE 13 MOL_ID: 3; SOURCE 14 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 15 ORGANISM_TAXID: 9606; SOURCE 16 EXPRESSION_SYSTEM: HOMO SAPIENS; SOURCE 17 EXPRESSION_SYSTEM_TAXID: 9606 KEYWDS NEUTRALIZING ANTIBODY, VIRAL FUSION PROTEIN, SARS-COV-2, VIRAL KEYWDS 2 PROTEIN-IMMUNE SYSTEM COMPLEX, VIRAL PROTEIN EXPDTA ELECTRON MICROSCOPY AUTHOR R.G.CASNER,L.SHAPIRO REVDAT 1 02-JUL-25 9CFH 0 JRNL AUTH R.G.CASNER JRNL TITL MORE SARS-COV-2 ANTIBODIES JRNL REF TO BE PUBLISHED JRNL REFN REMARK 2 REMARK 2 RESOLUTION. 3.10 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 SOFTWARE PACKAGES : LEGINON, PHENIX, CRYOSPARC REMARK 3 RECONSTRUCTION SCHEMA : NULL REMARK 3 REMARK 3 EM MAP-MODEL FITTING AND REFINEMENT REMARK 3 PDB ENTRY : NULL REMARK 3 REFINEMENT SPACE : NULL REMARK 3 REFINEMENT PROTOCOL : NULL REMARK 3 REFINEMENT TARGET : NULL REMARK 3 OVERALL ANISOTROPIC B VALUE : NULL REMARK 3 REMARK 3 FITTING PROCEDURE : NULL REMARK 3 REMARK 3 EM IMAGE RECONSTRUCTION STATISTICS REMARK 3 NOMINAL PIXEL SIZE (ANGSTROMS) : NULL REMARK 3 ACTUAL PIXEL SIZE (ANGSTROMS) : NULL REMARK 3 EFFECTIVE RESOLUTION (ANGSTROMS) : 3.100 REMARK 3 NUMBER OF PARTICLES : 102510 REMARK 3 CTF CORRECTION METHOD : PHASE FLIPPING AND AMPLITUDE REMARK 3 CORRECTION REMARK 3 REMARK 3 EM RECONSTRUCTION MAGNIFICATION CALIBRATION: NULL REMARK 3 REMARK 3 OTHER DETAILS: NULL REMARK 4 REMARK 4 9CFH COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 30-JUN-24. REMARK 100 THE DEPOSITION ID IS D_1000285413. REMARK 245 REMARK 245 EXPERIMENTAL DETAILS REMARK 245 RECONSTRUCTION METHOD : SINGLE PARTICLE REMARK 245 SPECIMEN TYPE : NULL REMARK 245 REMARK 245 ELECTRON MICROSCOPE SAMPLE REMARK 245 SAMPLE TYPE : PARTICLE REMARK 245 PARTICLE TYPE : POINT REMARK 245 NAME OF SAMPLE : ANTIBODY 19-77 IN COMPLEX WITH REMARK 245 SARS-COV-2 HK.3 RBD REMARK 245 SAMPLE CONCENTRATION (MG ML-1) : 1.00 REMARK 245 SAMPLE SUPPORT DETAILS : NULL REMARK 245 SAMPLE VITRIFICATION DETAILS : NULL REMARK 245 SAMPLE BUFFER : NULL REMARK 245 PH : 7.40 REMARK 245 SAMPLE DETAILS : NULL REMARK 245 REMARK 245 DATA ACQUISITION REMARK 245 DATE OF EXPERIMENT : NULL REMARK 245 NUMBER OF MICROGRAPHS-IMAGES : NULL REMARK 245 TEMPERATURE (KELVIN) : NULL REMARK 245 MICROSCOPE MODEL : FEI TITAN KRIOS REMARK 245 DETECTOR TYPE : GATAN K3 BIOQUANTUM (6K X REMARK 245 4K) REMARK 245 MINIMUM DEFOCUS (NM) : 800.00 REMARK 245 MAXIMUM DEFOCUS (NM) : 2000.00 REMARK 245 MINIMUM TILT ANGLE (DEGREES) : NULL REMARK 245 MAXIMUM TILT ANGLE (DEGREES) : NULL REMARK 245 NOMINAL CS : NULL REMARK 245 IMAGING MODE : BRIGHT FIELD REMARK 245 ELECTRON DOSE (ELECTRONS NM**-2) : 5800.00 REMARK 245 ILLUMINATION MODE : FLOOD BEAM REMARK 245 NOMINAL MAGNIFICATION : NULL REMARK 245 CALIBRATED MAGNIFICATION : NULL REMARK 245 SOURCE : FIELD EMISSION GUN REMARK 245 ACCELERATION VOLTAGE (KV) : 300 REMARK 245 IMAGING DETAILS : NULL REMARK 247 REMARK 247 ELECTRON MICROSCOPY REMARK 247 THE COORDINATES IN THIS ENTRY WERE GENERATED FROM ELECTRON REMARK 247 MICROSCOPY DATA. PROTEIN DATA BANK CONVENTIONS REQUIRE REMARK 247 THAT CRYST1 AND SCALE RECORDS BE INCLUDED, BUT THE VALUES REMARK 247 ON THESE RECORDS ARE MEANINGLESS EXCEPT FOR THE CALCULATION REMARK 247 OF THE STRUCTURE FACTORS. 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 SOFTWARE DETERMINED QUATERNARY STRUCTURE: TRIMERIC REMARK 350 SOFTWARE USED: PISA 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 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 ARG A 319 REMARK 465 VAL A 320 REMARK 465 GLN A 321 REMARK 465 PRO A 322 REMARK 465 THR A 323 REMARK 465 GLU A 324 REMARK 465 SER A 325 REMARK 465 ILE A 326 REMARK 465 VAL A 327 REMARK 465 ARG A 328 REMARK 465 PHE A 329 REMARK 465 PRO A 330 REMARK 465 ASN A 331 REMARK 465 ILE A 332 REMARK 465 THR A 333 REMARK 465 LYS A 528 REMARK 465 LYS A 529 REMARK 465 SER A 530 REMARK 465 THR A 531 REMARK 465 ASN A 532 REMARK 465 LEU A 533 REMARK 465 VAL A 534 REMARK 465 LYS A 535 REMARK 465 ASN A 536 REMARK 465 LYS A 537 REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: COVALENT BOND ANGLES 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,A3,1X,A1,I4,A1,3(1X,A4,2X),12X,F5.1) 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 ATM2 ATM3 REMARK 500 CYS A 432 CA - CB - SG ANGL. DEV. = 10.9 DEGREES 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 LYS A 378 113.90 -162.34 REMARK 500 TYR A 501 156.41 83.89 REMARK 500 LEU L 47 -60.15 -101.28 REMARK 500 ALA L 51 -9.26 72.63 REMARK 500 GLU L 68 -75.04 65.12 REMARK 500 ILE L 107 -50.53 -126.40 REMARK 500 REMARK 500 REMARK: NULL REMARK 900 REMARK 900 RELATED ENTRIES REMARK 900 RELATED ID: EMD-45546 RELATED DB: EMDB REMARK 900 CRYO-EM REFINEMENT OF ANTIBODY 19-77 R71V IN COMPLEX WITH SARS-COV- REMARK 900 2 JD.1.1 RBD DBREF 9CFH A 319 537 UNP P0DTC2 SPIKE_SARS2 319 537 DBREF 9CFH H 1 115 PDB 9CFH 9CFH 1 115 DBREF 9CFH L 1 108 PDB 9CFH 9CFH 1 108 SEQADV 9CFH HIS A 339 UNP P0DTC2 GLY 339 VARIANT SEQADV 9CFH THR A 346 UNP P0DTC2 ARG 346 VARIANT SEQADV 9CFH ILE A 368 UNP P0DTC2 LEU 368 VARIANT SEQADV 9CFH PHE A 371 UNP P0DTC2 SER 371 VARIANT SEQADV 9CFH PRO A 373 UNP P0DTC2 SER 373 VARIANT SEQADV 9CFH PHE A 375 UNP P0DTC2 SER 375 VARIANT SEQADV 9CFH ALA A 376 UNP P0DTC2 THR 376 VARIANT SEQADV 9CFH ASN A 405 UNP P0DTC2 ASP 405 VARIANT SEQADV 9CFH SER A 408 UNP P0DTC2 ARG 408 VARIANT SEQADV 9CFH ASN A 417 UNP P0DTC2 LYS 417 VARIANT SEQADV 9CFH LYS A 440 UNP P0DTC2 ASN 440 VARIANT SEQADV 9CFH PRO A 445 UNP P0DTC2 VAL 445 VARIANT SEQADV 9CFH SER A 446 UNP P0DTC2 GLY 446 VARIANT SEQADV 9CFH PHE A 455 UNP P0DTC2 LEU 455 CONFLICT SEQADV 9CFH LEU A 456 UNP P0DTC2 PHE 456 VARIANT SEQADV 9CFH LYS A 460 UNP P0DTC2 ASN 460 VARIANT SEQADV 9CFH VAL A 475 UNP P0DTC2 ALA 475 CONFLICT SEQADV 9CFH ASN A 477 UNP P0DTC2 SER 477 VARIANT SEQADV 9CFH LYS A 478 UNP P0DTC2 THR 478 VARIANT SEQADV 9CFH ALA A 484 UNP P0DTC2 GLU 484 VARIANT SEQADV 9CFH PRO A 486 UNP P0DTC2 PHE 486 VARIANT SEQADV 9CFH SER A 490 UNP P0DTC2 PHE 490 VARIANT SEQADV 9CFH ARG A 498 UNP P0DTC2 GLN 498 VARIANT SEQADV 9CFH TYR A 501 UNP P0DTC2 ASN 501 VARIANT SEQADV 9CFH HIS A 505 UNP P0DTC2 TYR 505 VARIANT SEQRES 1 A 219 ARG VAL GLN PRO THR GLU SER ILE VAL ARG PHE PRO ASN SEQRES 2 A 219 ILE THR ASN LEU CYS PRO PHE HIS GLU VAL PHE ASN ALA SEQRES 3 A 219 THR THR PHE ALA SER VAL TYR ALA TRP ASN ARG LYS ARG SEQRES 4 A 219 ILE SER ASN CYS VAL ALA ASP TYR SER VAL ILE TYR ASN SEQRES 5 A 219 PHE ALA PRO PHE PHE ALA PHE LYS CYS TYR GLY VAL SER SEQRES 6 A 219 PRO THR LYS LEU ASN ASP LEU CYS PHE THR ASN VAL TYR SEQRES 7 A 219 ALA ASP SER PHE VAL ILE ARG GLY ASN GLU VAL SER GLN SEQRES 8 A 219 ILE ALA PRO GLY GLN THR GLY ASN ILE ALA ASP TYR ASN SEQRES 9 A 219 TYR LYS LEU PRO ASP ASP PHE THR GLY CYS VAL ILE ALA SEQRES 10 A 219 TRP ASN SER ASN LYS LEU ASP SER LYS PRO SER GLY ASN SEQRES 11 A 219 TYR ASN TYR LEU TYR ARG PHE LEU ARG LYS SER LYS LEU SEQRES 12 A 219 LYS PRO PHE GLU ARG ASP ILE SER THR GLU ILE TYR GLN SEQRES 13 A 219 VAL GLY ASN LYS PRO CYS ASN GLY VAL ALA GLY PRO ASN SEQRES 14 A 219 CYS TYR SER PRO LEU GLN SER TYR GLY PHE ARG PRO THR SEQRES 15 A 219 TYR GLY VAL GLY HIS GLN PRO TYR ARG VAL VAL VAL LEU SEQRES 16 A 219 SER PHE GLU LEU LEU HIS ALA PRO ALA THR VAL CYS GLY SEQRES 17 A 219 PRO LYS LYS SER THR ASN LEU VAL LYS ASN LYS SEQRES 1 H 115 GLU VAL GLN VAL VAL GLU SER GLY GLY GLY LEU ILE GLN SEQRES 2 H 115 PRO GLY GLY SER LEU ARG LEU SER CYS THR ALA SER GLU SEQRES 3 H 115 LEU ILE ILE SER ARG ASN TYR MET SER TRP VAL ARG GLN SEQRES 4 H 115 ALA PRO GLY LYS GLY LEU GLU TRP LEU SER VAL ILE TYR SEQRES 5 H 115 PRO GLY GLY SER SER PHE TYR THR ASP SER LEU LYS GLY SEQRES 6 H 115 ARG PHE THR ILE SER VAL ASP ASN SER LYS ASN THR LEU SEQRES 7 H 115 TYR LEU GLN MET ASN ARG LEU GLY VAL GLU ASP THR ALA SEQRES 8 H 115 ILE TYR TYR CYS VAL ARG ASP ALA PRO SER GLU SER ASP SEQRES 9 H 115 TRP GLY GLN GLY THR LEU VAL THR VAL SER SER SEQRES 1 L 108 GLU ILE VAL LEU THR GLN SER PRO ALA THR LEU SER LEU SEQRES 2 L 108 PHE PRO GLY GLU ARG ALA THR LEU SER CYS ARG ALA SER SEQRES 3 L 108 GLN ASN ILE GLY HIS PHE LEU THR TRP TYR GLN GLN LYS SEQRES 4 L 108 PRO GLY GLN ALA PRO ARG LEU LEU ILE TYR ASP ALA SER SEQRES 5 L 108 ASN ARG ALA THR GLY VAL PRO ALA ARG PHE SER GLY SER SEQRES 6 L 108 GLY SER GLU THR GLU PHE THR LEU THR ILE SER SER LEU SEQRES 7 L 108 GLY PRO GLU ASP PHE ALA VAL TYR TYR CYS GLN GLU ARG SEQRES 8 L 108 SER ASP TRP PRO ARG GLY THR PHE GLY GLN GLY THR LYS SEQRES 9 L 108 VAL GLU ILE LYS HELIX 1 AA1 PRO A 337 ASN A 343 1 7 HELIX 2 AA2 SER A 349 TRP A 353 5 5 HELIX 3 AA3 TYR A 365 PHE A 371 1 7 HELIX 4 AA4 ASN A 405 ILE A 410 5 6 HELIX 5 AA5 GLY A 416 ASN A 422 1 7 HELIX 6 AA6 SER A 438 SER A 443 1 6 HELIX 7 AA7 ILE H 28 ASN H 32 5 5 HELIX 8 AA8 ASP H 61 LYS H 64 5 4 HELIX 9 AA9 GLY H 86 THR H 90 5 5 HELIX 10 AB1 GLY L 79 PHE L 83 5 5 SHEET 1 AA1 5 ASN A 354 ILE A 358 0 SHEET 2 AA1 5 ASN A 394 ARG A 403 -1 O VAL A 395 N ILE A 358 SHEET 3 AA1 5 PRO A 507 GLU A 516 -1 O SER A 514 N TYR A 396 SHEET 4 AA1 5 GLY A 431 ASN A 437 -1 N ILE A 434 O VAL A 511 SHEET 5 AA1 5 ALA A 376 TYR A 380 -1 N ALA A 376 O ALA A 435 SHEET 1 AA2 3 CYS A 361 VAL A 362 0 SHEET 2 AA2 3 VAL A 524 CYS A 525 1 O CYS A 525 N CYS A 361 SHEET 3 AA2 3 CYS A 391 PHE A 392 -1 N PHE A 392 O VAL A 524 SHEET 1 AA3 2 LEU A 452 ARG A 454 0 SHEET 2 AA3 2 LEU A 492 SER A 494 -1 O GLN A 493 N TYR A 453 SHEET 1 AA4 2 TYR A 473 GLN A 474 0 SHEET 2 AA4 2 CYS A 488 TYR A 489 -1 O TYR A 489 N TYR A 473 SHEET 1 AA5 4 VAL H 4 SER H 7 0 SHEET 2 AA5 4 SER H 17 ALA H 24 -1 O THR H 23 N VAL H 5 SHEET 3 AA5 4 THR H 77 ASN H 83 -1 O LEU H 78 N CYS H 22 SHEET 4 AA5 4 PHE H 67 ASP H 72 -1 N SER H 70 O TYR H 79 SHEET 1 AA6 6 GLY H 10 ILE H 12 0 SHEET 2 AA6 6 THR H 109 VAL H 113 1 O LEU H 110 N GLY H 10 SHEET 3 AA6 6 ALA H 91 ASP H 98 -1 N TYR H 93 O THR H 109 SHEET 4 AA6 6 TYR H 33 GLN H 39 -1 N VAL H 37 O TYR H 94 SHEET 5 AA6 6 LEU H 45 ILE H 51 -1 O LEU H 48 N TRP H 36 SHEET 6 AA6 6 SER H 57 TYR H 59 -1 O PHE H 58 N VAL H 50 SHEET 1 AA7 4 GLY H 10 ILE H 12 0 SHEET 2 AA7 4 THR H 109 VAL H 113 1 O LEU H 110 N GLY H 10 SHEET 3 AA7 4 ALA H 91 ASP H 98 -1 N TYR H 93 O THR H 109 SHEET 4 AA7 4 SER H 103 TRP H 105 -1 O ASP H 104 N ARG H 97 SHEET 1 AA8 4 LEU L 4 SER L 7 0 SHEET 2 AA8 4 ALA L 19 ALA L 25 -1 O ARG L 24 N THR L 5 SHEET 3 AA8 4 GLU L 70 ILE L 75 -1 O LEU L 73 N LEU L 21 SHEET 4 AA8 4 PHE L 62 SER L 67 -1 N SER L 63 O THR L 74 SHEET 1 AA9 6 THR L 10 LEU L 11 0 SHEET 2 AA9 6 THR L 103 VAL L 105 1 O LYS L 104 N LEU L 11 SHEET 3 AA9 6 VAL L 85 GLU L 90 -1 N TYR L 86 O THR L 103 SHEET 4 AA9 6 LEU L 33 GLN L 38 -1 N THR L 34 O GLN L 89 SHEET 5 AA9 6 ARG L 45 TYR L 49 -1 O ARG L 45 N GLN L 37 SHEET 6 AA9 6 ASN L 53 ARG L 54 -1 O ASN L 53 N TYR L 49 SSBOND 1 CYS A 336 CYS A 361 1555 1555 2.03 SSBOND 2 CYS A 379 CYS A 432 1555 1555 2.05 SSBOND 3 CYS A 391 CYS A 525 1555 1555 2.03 SSBOND 4 CYS A 480 CYS A 488 1555 1555 2.04 SSBOND 5 CYS H 22 CYS H 95 1555 1555 2.03 SSBOND 6 CYS L 23 CYS L 88 1555 1555 2.04 CISPEP 1 SER L 7 PRO L 8 0 -2.32 CISPEP 2 TRP L 94 PRO L 95 0 2.96 CRYST1 1.000 1.000 1.000 90.00 90.00 90.00 P 1 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 1.000000 0.000000 0.000000 0.00000 SCALE2 0.000000 1.000000 0.000000 0.00000 SCALE3 0.000000 0.000000 1.000000 0.00000