HEADER OXIDOREDUCTASE 11-DEC-24 9HO5 TITLE CRYSTAL STRUCTURE OF THE HUMAN FRATAXIN PROTEIN IN COMPLEX WITH A TITLE 2 TAILORED CAMELID NANOBODY 4A7 COMPND MOL_ID: 1; COMPND 2 MOLECULE: FRATAXIN MATURE FORM; COMPND 3 CHAIN: A; COMPND 4 SYNONYM: FRATAXIN(81-210),M81-FXN; COMPND 5 ENGINEERED: YES; COMPND 6 MOL_ID: 2; COMPND 7 MOLECULE: CAMELID NANOBODY 4A7; COMPND 8 CHAIN: C; COMPND 9 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: FXN, FRDA, X25; SOURCE 6 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 7 EXPRESSION_SYSTEM_TAXID: 562; SOURCE 8 MOL_ID: 2; SOURCE 9 ORGANISM_SCIENTIFIC: LAMA GLAMA; SOURCE 10 ORGANISM_COMMON: LLAMA; SOURCE 11 ORGANISM_TAXID: 9844; SOURCE 12 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 13 EXPRESSION_SYSTEM_TAXID: 562 KEYWDS COMPLEX, NANOBODY, MITOCHONDRIAL REGULATION, FRIEDREICH ATAXIA, KEYWDS 2 OXIDOREDUCTASE EXPDTA X-RAY DIFFRACTION AUTHOR A.GARAY-ALVAREZ,R.MOLINA,J.A.HERMOSO REVDAT 1 24-DEC-25 9HO5 0 JRNL AUTH M.F.PIGNATARO,N.B.FRENANDEZ,M.F.PAVAN,A.GARAY-ALVAREZ, JRNL AUTH 2 R.MOLINA,J.GROSSI,M.NOGUERA,A.VILA,A.GARCIA,H.GENTILI, JRNL AUTH 3 N.A.RODRIGUEZ,M.ARAN,J.A.HERMOSO,L.I.IBANEZ,J.SANTOS JRNL TITL FRATAXIN STABILITY AND FUNCTION MODULATION BY NANOBODY JRNL TITL 2 INTERACTION JRNL REF TO BE PUBLISHED JRNL REFN REMARK 2 REMARK 2 RESOLUTION. 1.50 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : PHENIX 1.21RC1_4958 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) : 1.50 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 55.71 REMARK 3 MIN(FOBS/SIGMA_FOBS) : 1.400 REMARK 3 COMPLETENESS FOR RANGE (%) : 100.0 REMARK 3 NUMBER OF REFLECTIONS : 34017 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 R VALUE (WORKING + TEST SET) : 0.188 REMARK 3 R VALUE (WORKING SET) : 0.187 REMARK 3 FREE R VALUE : 0.210 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 5.030 REMARK 3 FREE R VALUE TEST SET COUNT : 1710 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 55.7100 - 3.4300 1.00 2856 176 0.1702 0.1826 REMARK 3 2 3.4300 - 2.7300 1.00 2758 142 0.1937 0.2271 REMARK 3 3 2.7300 - 2.3800 1.00 2712 138 0.2032 0.2361 REMARK 3 4 2.3800 - 2.1600 1.00 2705 141 0.1929 0.2123 REMARK 3 5 2.1600 - 2.0100 1.00 2688 141 0.1870 0.1982 REMARK 3 6 2.0100 - 1.8900 1.00 2693 120 0.1912 0.2253 REMARK 3 7 1.8900 - 1.8000 1.00 2668 134 0.1928 0.2422 REMARK 3 8 1.8000 - 1.7200 1.00 2672 135 0.1872 0.2128 REMARK 3 9 1.7200 - 1.6500 1.00 2631 158 0.1879 0.2241 REMARK 3 10 1.6500 - 1.5900 1.00 2651 146 0.1884 0.2451 REMARK 3 11 1.5900 - 1.5400 1.00 2607 157 0.1899 0.2261 REMARK 3 12 1.5400 - 1.5000 1.00 2666 122 0.1897 0.2007 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) : 0.153 REMARK 3 PHASE ERROR (DEGREES, MAXIMUM-LIKELIHOOD BASED) : 19.178 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : 18.24 REMARK 3 MEAN B VALUE (OVERALL, A**2) : 20.81 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 : 0.006 1890 REMARK 3 ANGLE : 0.845 2567 REMARK 3 CHIRALITY : 0.055 283 REMARK 3 PLANARITY : 0.006 333 REMARK 3 DIHEDRAL : 17.973 680 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): 15.3357 -4.3915 -12.4102 REMARK 3 T TENSOR REMARK 3 T11: 0.1296 T22: 0.1071 REMARK 3 T33: 0.1420 T12: 0.0020 REMARK 3 T13: 0.0081 T23: -0.0148 REMARK 3 L TENSOR REMARK 3 L11: 0.5813 L22: 0.2067 REMARK 3 L33: 1.0164 L12: 0.2075 REMARK 3 L13: 0.3385 L23: 0.1323 REMARK 3 S TENSOR REMARK 3 S11: -0.0341 S12: 0.0327 S13: 0.0504 REMARK 3 S21: -0.0514 S22: -0.0030 S23: -0.0090 REMARK 3 S31: 0.0365 S32: 0.0186 S33: -0.0001 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 9HO5 COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY PDBE ON 11-DEC-24. REMARK 100 THE DEPOSITION ID IS D_1292143925. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 16-SEP-23 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 : 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 : AUTOPROC REMARK 200 DATA SCALING SOFTWARE : AIMLESS REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 47731 REMARK 200 RESOLUTION RANGE HIGH (A) : 1.246 REMARK 200 RESOLUTION RANGE LOW (A) : 55.713 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : NULL REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 89.3 REMARK 200 DATA REDUNDANCY : 10.70 REMARK 200 R MERGE (I) : NULL REMARK 200 R SYM (I) : NULL REMARK 200 FOR THE DATA SET : 23.6000 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 1.25 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 1.33 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: PHASER REMARK 200 STARTING MODEL: NULL REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 31.19 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 1.79 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 0.2M SODIUM THIOCYANATE PH 6.9 20% PEG REMARK 280 3350, VAPOR DIFFUSION, SITTING DROP, TEMPERATURE 291K 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 19.26150 REMARK 290 SMTRY2 2 0.000000 -1.000000 0.000000 0.00000 REMARK 290 SMTRY3 2 0.000000 0.000000 1.000000 55.71250 REMARK 290 SMTRY1 3 -1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 3 0.000000 1.000000 0.000000 24.07550 REMARK 290 SMTRY3 3 0.000000 0.000000 -1.000000 55.71250 REMARK 290 SMTRY1 4 1.000000 0.000000 0.000000 19.26150 REMARK 290 SMTRY2 4 0.000000 -1.000000 0.000000 24.07550 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 APPLY THE FOLLOWING TO CHAINS: A, C 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 A 0 REMARK 465 SER C 27 REMARK 465 VAL C 28 REMARK 465 PHE C 29 REMARK 465 SER C 30 REMARK 465 VAL C 31 REMARK 465 TYR C 124 REMARK 465 PRO C 125 REMARK 465 TYR C 126 REMARK 465 ASP C 127 REMARK 465 VAL C 128 REMARK 465 PRO C 129 REMARK 465 ASP C 130 REMARK 465 TYR C 131 REMARK 465 GLY C 132 REMARK 465 SER C 133 REMARK 465 HIS C 134 REMARK 465 HIS C 135 REMARK 465 HIS C 136 REMARK 465 HIS C 137 REMARK 465 HIS C 138 REMARK 465 HIS C 139 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 TRP C 32 CG CD1 CD2 NE1 CE2 CE3 CZ2 REMARK 470 TRP C 32 CZ3 CH2 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 NH1 ARG C 43 O HOH C 201 2.17 REMARK 500 O HOH A 276 O HOH C 261 2.19 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 120 73.75 -64.52 REMARK 500 ALA C 91 165.51 178.10 REMARK 500 REMARK 500 REMARK: NULL REMARK 900 REMARK 900 RELATED ENTRIES REMARK 900 RELATED ID: 9HO4 RELATED DB: PDB DBREF 9HO5 A 1 121 UNP Q16595 FRDA_HUMAN 90 210 DBREF 9HO5 C 1 139 PDB 9HO5 9HO5 1 139 SEQADV 9HO5 MET A 0 UNP Q16595 INITIATING METHIONINE SEQRES 1 A 122 MET LEU ASP GLU THR THR TYR GLU ARG LEU ALA GLU GLU SEQRES 2 A 122 THR LEU ASP SER LEU ALA GLU PHE PHE GLU ASP LEU ALA SEQRES 3 A 122 ASP LYS PRO TYR THR PHE GLU ASP TYR ASP VAL SER PHE SEQRES 4 A 122 GLY SER GLY VAL LEU THR VAL LYS LEU GLY GLY ASP LEU SEQRES 5 A 122 GLY THR TYR VAL ILE ASN LYS GLN THR PRO ASN LYS GLN SEQRES 6 A 122 ILE TRP LEU SER SER PRO SER SER GLY PRO LYS ARG TYR SEQRES 7 A 122 ASP TRP THR GLY LYS ASN TRP VAL TYR SER HIS ASP GLY SEQRES 8 A 122 VAL SER LEU HIS GLU LEU LEU ALA ALA GLU LEU THR LYS SEQRES 9 A 122 ALA LEU LYS THR LYS LEU ASP LEU SER SER LEU ALA TYR SEQRES 10 A 122 SER GLY LYS ASP ALA SEQRES 1 C 139 GLN VAL GLN LEU GLN GLU SER GLY GLY GLY SER VAL GLN SEQRES 2 C 139 PRO GLY GLY SER LEU ARG LEU SER CYS VAL ALA ARG GLY SEQRES 3 C 139 SER VAL PHE SER VAL TRP ASP MET GLY TRP TYR ARG GLN SEQRES 4 C 139 VAL PRO GLY ARG GLY ARG HIS LEU VAL ALA ARG LEU ASN SEQRES 5 C 139 ASP ASP GLY SER THR ASN TYR ALA ASP SER VAL LYS GLY SEQRES 6 C 139 ARG PHE THR ILE SER ARG ASP ASN ALA LYS ASN THR LEU SEQRES 7 C 139 TYR LEU GLN MET ASN SER LEU LYS PRO GLU ASP THR ALA SEQRES 8 C 139 VAL TYR TYR CYS ASN ALA VAL PRO PRO ILE VAL THR VAL SEQRES 9 C 139 ASN ALA THR ASP TYR TRP GLY GLN GLY THR GLN VAL THR SEQRES 10 C 139 VAL SER SER ALA ALA ALA TYR PRO TYR ASP VAL PRO ASP SEQRES 11 C 139 TYR GLY SER HIS HIS HIS HIS HIS HIS FORMUL 3 HOH *196(H2 O) HELIX 1 AA1 ASP A 2 ALA A 25 1 24 HELIX 2 AA2 SER A 92 LYS A 106 1 15 HELIX 3 AA3 LYS C 86 THR C 90 5 5 SHEET 1 AA1 6 ASP A 35 GLY A 39 0 SHEET 2 AA1 6 VAL A 42 LYS A 46 -1 O THR A 44 N SER A 37 SHEET 3 AA1 6 THR A 53 GLN A 59 -1 O ILE A 56 N LEU A 43 SHEET 4 AA1 6 GLN A 64 SER A 69 -1 O TRP A 66 N ASN A 57 SHEET 5 AA1 6 GLY A 73 TRP A 79 -1 O LYS A 75 N LEU A 67 SHEET 6 AA1 6 TRP A 84 TYR A 86 -1 O VAL A 85 N ASP A 78 SHEET 1 AA2 4 GLN C 3 SER C 7 0 SHEET 2 AA2 4 LEU C 18 ARG C 25 -1 O SER C 21 N SER C 7 SHEET 3 AA2 4 THR C 77 MET C 82 -1 O MET C 82 N LEU C 18 SHEET 4 AA2 4 PHE C 67 ASP C 72 -1 N SER C 70 O TYR C 79 SHEET 1 AA3 6 GLY C 10 VAL C 12 0 SHEET 2 AA3 6 THR C 114 VAL C 118 1 O THR C 117 N VAL C 12 SHEET 3 AA3 6 ALA C 91 ALA C 97 -1 N TYR C 93 O THR C 114 SHEET 4 AA3 6 MET C 34 GLN C 39 -1 N TYR C 37 O TYR C 94 SHEET 5 AA3 6 ARG C 45 LEU C 51 -1 O LEU C 51 N MET C 34 SHEET 6 AA3 6 THR C 57 TYR C 59 -1 O ASN C 58 N ARG C 50 SSBOND 1 CYS C 22 CYS C 95 1555 1555 2.03 CISPEP 1 VAL C 98 PRO C 99 0 -6.90 CRYST1 38.523 48.151 111.425 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 0.025959 0.000000 0.000000 0.00000 SCALE2 0.000000 0.020768 0.000000 0.00000 SCALE3 0.000000 0.000000 0.008975 0.00000