HEADER VIRAL PROTEIN 01-APR-24 9B9E TITLE PREFUSION F GLYCOPROTEIN ECTODOMAIN OF NIPAH VIRUS ECTODOMAIN IN TITLE 2 COMPLEX WITH DS90 NANOBODY COMPND MOL_ID: 1; COMPND 2 MOLECULE: FUSION GLYCOPROTEIN F0; COMPND 3 CHAIN: A, B, C; COMPND 4 FRAGMENT: ECTODOMAIN; COMPND 5 SYNONYM: PROTEIN F; COMPND 6 ENGINEERED: YES; COMPND 7 MOL_ID: 2; COMPND 8 MOLECULE: DS90 NANOBODY; COMPND 9 CHAIN: D, E, F; COMPND 10 ENGINEERED: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: HENIPAVIRUS NIPAHENSE; SOURCE 3 ORGANISM_TAXID: 3052225; SOURCE 4 EXPRESSION_SYSTEM: CRICETULUS GRISEUS; SOURCE 5 EXPRESSION_SYSTEM_TAXID: 10029; SOURCE 6 MOL_ID: 2; SOURCE 7 ORGANISM_SCIENTIFIC: VICUGNA PACOS; SOURCE 8 ORGANISM_TAXID: 30538; SOURCE 9 EXPRESSION_SYSTEM: CRICETULUS GRISEUS; SOURCE 10 EXPRESSION_SYSTEM_TAXID: 10029 KEYWDS F ECTODOMAIN, PREFUSION, VIRAL PROTEIN, NIPAH VIRUS EXPDTA ELECTRON MICROSCOPY AUTHOR Y.S.LOW,A.ISAACS,N.MODHIRAN,D.WATTERSON REVDAT 1 04-JUN-25 9B9E 0 JRNL AUTH Y.S.LOW,A.ISAACS,N.MODHIRAN,D.WATTERSON JRNL TITL PREFUSION F GLYCOPROTEIN ECTODOMAIN OF NIPAH VIRUS JRNL TITL 2 ECTODOMAIN IN COMPLEX WITH DS90 NANOBODY JRNL REF TO BE PUBLISHED JRNL REFN REMARK 2 REMARK 2 RESOLUTION. 3.66 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 SOFTWARE PACKAGES : CRYOSPARC, SERIALEM, CRYOSPARC, ISOLDE, REMARK 3 COOT, CRYOSPARC, CRYOSPARC, CRYOSPARC, REMARK 3 CRYOSPARC, PHENIX REMARK 3 RECONSTRUCTION SCHEMA : NULL REMARK 3 REMARK 3 EM MAP-MODEL FITTING AND REFINEMENT REMARK 3 PDB ENTRY : NULL REMARK 3 REFINEMENT SPACE : REAL REMARK 3 REFINEMENT PROTOCOL : FLEXIBLE FIT REMARK 3 REFINEMENT TARGET : MOLPROBITY REMARK 3 OVERALL ANISOTROPIC B VALUE : 99.700 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) : 3.660 REMARK 3 NUMBER OF PARTICLES : 200640 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 9B9E COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 02-APR-24. REMARK 100 THE DEPOSITION ID IS D_1000282783. 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 : PREFUSION F GLYCOPROTEIN REMARK 245 ECTODOMAIN OF NIPAH VIRUS REMARK 245 ECTODOMAIN IN COMPLEX WITH DS90 REMARK 245 NANOBODY REMARK 245 SAMPLE CONCENTRATION (MG ML-1) : 0.04 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 : JEOL CRYO ARM 300 REMARK 245 DETECTOR TYPE : GATAN K3 (6K X 4K) REMARK 245 MINIMUM DEFOCUS (NM) : 5000.00 REMARK 245 MAXIMUM DEFOCUS (NM) : 25000.00 REMARK 245 MINIMUM TILT ANGLE (DEGREES) : NULL REMARK 245 MAXIMUM TILT ANGLE (DEGREES) : NULL REMARK 245 NOMINAL CS : 2.70 REMARK 245 IMAGING MODE : BRIGHT FIELD REMARK 245 ELECTRON DOSE (ELECTRONS NM**-2) : 8750.00 REMARK 245 ILLUMINATION MODE : OTHER REMARK 245 NOMINAL MAGNIFICATION : 60000 REMARK 245 CALIBRATED MAGNIFICATION : 60000 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: HEXAMERIC REMARK 350 SOFTWARE DETERMINED QUATERNARY STRUCTURE: HEXAMERIC REMARK 350 SOFTWARE USED: PISA REMARK 350 APPLY THE FOLLOWING TO CHAINS: A, B, C, D, E, F 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 THR A 483 REMARK 465 THR B 483 REMARK 465 THR C 483 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 B 394 CA - CB - SG ANGL. DEV. = 7.8 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 ASN A 51 61.93 37.88 REMARK 500 MET A 68 32.05 -143.49 REMARK 500 ASN A 155 27.64 -148.36 REMARK 500 LEU A 218 60.75 -105.79 REMARK 500 ASP A 220 56.56 -119.27 REMARK 500 SER A 273 -4.82 -140.72 REMARK 500 ASP A 304 -122.73 48.92 REMARK 500 CYS A 387 -3.65 -58.65 REMARK 500 ASN B 51 63.17 32.55 REMARK 500 MET B 68 33.67 -142.87 REMARK 500 ASN B 155 25.53 -146.26 REMARK 500 LEU B 218 63.12 -104.78 REMARK 500 ASP B 304 -125.05 52.19 REMARK 500 CYS B 387 -8.69 -55.20 REMARK 500 ASN C 51 64.11 32.62 REMARK 500 MET C 68 33.55 -142.81 REMARK 500 ASN C 155 25.70 -146.62 REMARK 500 LEU C 218 62.76 -104.09 REMARK 500 ASP C 304 -125.12 52.30 REMARK 500 CYS C 387 -9.00 -54.48 REMARK 500 SER D 29 3.13 -69.56 REMARK 500 REMARK 500 REMARK: NULL REMARK 900 REMARK 900 RELATED ENTRIES REMARK 900 RELATED ID: EMD-44380 RELATED DB: EMDB REMARK 900 PREFUSION F GLYCOPROTEIN ECTODOMAIN OF NIPAH VIRUS ECTODOMAIN IN REMARK 900 COMPLEX WITH DS90 NANOBODY DBREF 9B9E A 27 483 UNP Q9IH63 FUS_NIPAV 27 483 DBREF 9B9E B 27 483 UNP Q9IH63 FUS_NIPAV 27 483 DBREF 9B9E C 27 483 UNP Q9IH63 FUS_NIPAV 27 483 DBREF 9B9E D 1 124 PDB 9B9E 9B9E 1 124 DBREF 9B9E E 1 124 PDB 9B9E 9B9E 1 124 DBREF 9B9E F 1 124 PDB 9B9E 9B9E 1 124 SEQRES 1 A 457 ILE LEU HIS TYR GLU LYS LEU SER LYS ILE GLY LEU VAL SEQRES 2 A 457 LYS GLY VAL THR ARG LYS TYR LYS ILE LYS SER ASN PRO SEQRES 3 A 457 LEU THR LYS ASP ILE VAL ILE LYS MET ILE PRO ASN VAL SEQRES 4 A 457 SER ASN MET SER GLN CYS THR GLY SER VAL MET GLU ASN SEQRES 5 A 457 TYR LYS THR ARG LEU ASN GLY ILE LEU THR PRO ILE LYS SEQRES 6 A 457 GLY ALA LEU GLU ILE TYR LYS ASN ASN THR HIS ASP LEU SEQRES 7 A 457 VAL GLY ASP VAL ARG LEU ALA GLY VAL ILE MET ALA GLY SEQRES 8 A 457 VAL ALA ILE GLY ILE ALA THR ALA ALA GLN ILE THR ALA SEQRES 9 A 457 GLY VAL ALA LEU TYR GLU ALA MET LYS ASN ALA ASP ASN SEQRES 10 A 457 ILE ASN LYS LEU LYS SER SER ILE GLU SER THR ASN GLU SEQRES 11 A 457 ALA VAL VAL LYS LEU GLN GLU THR ALA GLU LYS THR VAL SEQRES 12 A 457 TYR VAL LEU THR ALA LEU GLN ASP TYR ILE ASN THR ASN SEQRES 13 A 457 LEU VAL PRO THR ILE ASP LYS ILE SER CYS LYS GLN THR SEQRES 14 A 457 GLU LEU SER LEU ASP LEU ALA LEU SER LYS TYR LEU SER SEQRES 15 A 457 ASP LEU LEU PHE VAL PHE GLY PRO ASN LEU GLN ASP PRO SEQRES 16 A 457 VAL SER ASN SER MET THR ILE GLN ALA ILE SER GLN ALA SEQRES 17 A 457 PHE GLY GLY ASN TYR GLU THR LEU LEU ARG THR LEU GLY SEQRES 18 A 457 TYR ALA THR GLU ASP PHE ASP ASP LEU LEU GLU SER ASP SEQRES 19 A 457 SER ILE THR GLY GLN ILE ILE TYR VAL ASP LEU SER SER SEQRES 20 A 457 TYR TYR ILE ILE VAL ARG VAL TYR PHE PRO ILE LEU THR SEQRES 21 A 457 GLU ILE GLN GLN ALA TYR ILE GLN GLU LEU LEU PRO VAL SEQRES 22 A 457 SER PHE ASN ASN ASP ASN SER GLU TRP ILE SER ILE VAL SEQRES 23 A 457 PRO ASN PHE ILE LEU VAL ARG ASN THR LEU ILE SER ASN SEQRES 24 A 457 ILE GLU ILE GLY PHE CYS LEU ILE THR LYS ARG SER VAL SEQRES 25 A 457 ILE CYS ASN GLN ASP TYR ALA THR PRO MET THR ASN ASN SEQRES 26 A 457 MET ARG GLU CYS LEU THR GLY SER THR GLU LYS CYS PRO SEQRES 27 A 457 ARG GLU LEU VAL VAL SER SER HIS VAL PRO ARG PHE ALA SEQRES 28 A 457 LEU SER ASN GLY VAL LEU PHE ALA ASN CYS ILE SER VAL SEQRES 29 A 457 THR CYS GLN CYS GLN THR THR GLY ARG ALA ILE SER GLN SEQRES 30 A 457 SER GLY GLU GLN THR LEU LEU MET ILE ASP ASN THR THR SEQRES 31 A 457 CYS PRO THR ALA VAL LEU GLY ASN VAL ILE ILE SER LEU SEQRES 32 A 457 GLY LYS TYR LEU GLY SER VAL ASN TYR ASN SER GLU GLY SEQRES 33 A 457 ILE ALA ILE GLY PRO PRO VAL PHE THR ASP LYS VAL ASP SEQRES 34 A 457 ILE SER SER GLN ILE SER SER MET ASN GLN SER LEU GLN SEQRES 35 A 457 GLN SER LYS ASP TYR ILE LYS GLU ALA GLN ARG LEU LEU SEQRES 36 A 457 ASP THR SEQRES 1 B 457 ILE LEU HIS TYR GLU LYS LEU SER LYS ILE GLY LEU VAL SEQRES 2 B 457 LYS GLY VAL THR ARG LYS TYR LYS ILE LYS SER ASN PRO SEQRES 3 B 457 LEU THR LYS ASP ILE VAL ILE LYS MET ILE PRO ASN VAL SEQRES 4 B 457 SER ASN MET SER GLN CYS THR GLY SER VAL MET GLU ASN SEQRES 5 B 457 TYR LYS THR ARG LEU ASN GLY ILE LEU THR PRO ILE LYS SEQRES 6 B 457 GLY ALA LEU GLU ILE TYR LYS ASN ASN THR HIS ASP LEU SEQRES 7 B 457 VAL GLY ASP VAL ARG LEU ALA GLY VAL ILE MET ALA GLY SEQRES 8 B 457 VAL ALA ILE GLY ILE ALA THR ALA ALA GLN ILE THR ALA SEQRES 9 B 457 GLY VAL ALA LEU TYR GLU ALA MET LYS ASN ALA ASP ASN SEQRES 10 B 457 ILE ASN LYS LEU LYS SER SER ILE GLU SER THR ASN GLU SEQRES 11 B 457 ALA VAL VAL LYS LEU GLN GLU THR ALA GLU LYS THR VAL SEQRES 12 B 457 TYR VAL LEU THR ALA LEU GLN ASP TYR ILE ASN THR ASN SEQRES 13 B 457 LEU VAL PRO THR ILE ASP LYS ILE SER CYS LYS GLN THR SEQRES 14 B 457 GLU LEU SER LEU ASP LEU ALA LEU SER LYS TYR LEU SER SEQRES 15 B 457 ASP LEU LEU PHE VAL PHE GLY PRO ASN LEU GLN ASP PRO SEQRES 16 B 457 VAL SER ASN SER MET THR ILE GLN ALA ILE SER GLN ALA SEQRES 17 B 457 PHE GLY GLY ASN TYR GLU THR LEU LEU ARG THR LEU GLY SEQRES 18 B 457 TYR ALA THR GLU ASP PHE ASP ASP LEU LEU GLU SER ASP SEQRES 19 B 457 SER ILE THR GLY GLN ILE ILE TYR VAL ASP LEU SER SER SEQRES 20 B 457 TYR TYR ILE ILE VAL ARG VAL TYR PHE PRO ILE LEU THR SEQRES 21 B 457 GLU ILE GLN GLN ALA TYR ILE GLN GLU LEU LEU PRO VAL SEQRES 22 B 457 SER PHE ASN ASN ASP ASN SER GLU TRP ILE SER ILE VAL SEQRES 23 B 457 PRO ASN PHE ILE LEU VAL ARG ASN THR LEU ILE SER ASN SEQRES 24 B 457 ILE GLU ILE GLY PHE CYS LEU ILE THR LYS ARG SER VAL SEQRES 25 B 457 ILE CYS ASN GLN ASP TYR ALA THR PRO MET THR ASN ASN SEQRES 26 B 457 MET ARG GLU CYS LEU THR GLY SER THR GLU LYS CYS PRO SEQRES 27 B 457 ARG GLU LEU VAL VAL SER SER HIS VAL PRO ARG PHE ALA SEQRES 28 B 457 LEU SER ASN GLY VAL LEU PHE ALA ASN CYS ILE SER VAL SEQRES 29 B 457 THR CYS GLN CYS GLN THR THR GLY ARG ALA ILE SER GLN SEQRES 30 B 457 SER GLY GLU GLN THR LEU LEU MET ILE ASP ASN THR THR SEQRES 31 B 457 CYS PRO THR ALA VAL LEU GLY ASN VAL ILE ILE SER LEU SEQRES 32 B 457 GLY LYS TYR LEU GLY SER VAL ASN TYR ASN SER GLU GLY SEQRES 33 B 457 ILE ALA ILE GLY PRO PRO VAL PHE THR ASP LYS VAL ASP SEQRES 34 B 457 ILE SER SER GLN ILE SER SER MET ASN GLN SER LEU GLN SEQRES 35 B 457 GLN SER LYS ASP TYR ILE LYS GLU ALA GLN ARG LEU LEU SEQRES 36 B 457 ASP THR SEQRES 1 C 457 ILE LEU HIS TYR GLU LYS LEU SER LYS ILE GLY LEU VAL SEQRES 2 C 457 LYS GLY VAL THR ARG LYS TYR LYS ILE LYS SER ASN PRO SEQRES 3 C 457 LEU THR LYS ASP ILE VAL ILE LYS MET ILE PRO ASN VAL SEQRES 4 C 457 SER ASN MET SER GLN CYS THR GLY SER VAL MET GLU ASN SEQRES 5 C 457 TYR LYS THR ARG LEU ASN GLY ILE LEU THR PRO ILE LYS SEQRES 6 C 457 GLY ALA LEU GLU ILE TYR LYS ASN ASN THR HIS ASP LEU SEQRES 7 C 457 VAL GLY ASP VAL ARG LEU ALA GLY VAL ILE MET ALA GLY SEQRES 8 C 457 VAL ALA ILE GLY ILE ALA THR ALA ALA GLN ILE THR ALA SEQRES 9 C 457 GLY VAL ALA LEU TYR GLU ALA MET LYS ASN ALA ASP ASN SEQRES 10 C 457 ILE ASN LYS LEU LYS SER SER ILE GLU SER THR ASN GLU SEQRES 11 C 457 ALA VAL VAL LYS LEU GLN GLU THR ALA GLU LYS THR VAL SEQRES 12 C 457 TYR VAL LEU THR ALA LEU GLN ASP TYR ILE ASN THR ASN SEQRES 13 C 457 LEU VAL PRO THR ILE ASP LYS ILE SER CYS LYS GLN THR SEQRES 14 C 457 GLU LEU SER LEU ASP LEU ALA LEU SER LYS TYR LEU SER SEQRES 15 C 457 ASP LEU LEU PHE VAL PHE GLY PRO ASN LEU GLN ASP PRO SEQRES 16 C 457 VAL SER ASN SER MET THR ILE GLN ALA ILE SER GLN ALA SEQRES 17 C 457 PHE GLY GLY ASN TYR GLU THR LEU LEU ARG THR LEU GLY SEQRES 18 C 457 TYR ALA THR GLU ASP PHE ASP ASP LEU LEU GLU SER ASP SEQRES 19 C 457 SER ILE THR GLY GLN ILE ILE TYR VAL ASP LEU SER SER SEQRES 20 C 457 TYR TYR ILE ILE VAL ARG VAL TYR PHE PRO ILE LEU THR SEQRES 21 C 457 GLU ILE GLN GLN ALA TYR ILE GLN GLU LEU LEU PRO VAL SEQRES 22 C 457 SER PHE ASN ASN ASP ASN SER GLU TRP ILE SER ILE VAL SEQRES 23 C 457 PRO ASN PHE ILE LEU VAL ARG ASN THR LEU ILE SER ASN SEQRES 24 C 457 ILE GLU ILE GLY PHE CYS LEU ILE THR LYS ARG SER VAL SEQRES 25 C 457 ILE CYS ASN GLN ASP TYR ALA THR PRO MET THR ASN ASN SEQRES 26 C 457 MET ARG GLU CYS LEU THR GLY SER THR GLU LYS CYS PRO SEQRES 27 C 457 ARG GLU LEU VAL VAL SER SER HIS VAL PRO ARG PHE ALA SEQRES 28 C 457 LEU SER ASN GLY VAL LEU PHE ALA ASN CYS ILE SER VAL SEQRES 29 C 457 THR CYS GLN CYS GLN THR THR GLY ARG ALA ILE SER GLN SEQRES 30 C 457 SER GLY GLU GLN THR LEU LEU MET ILE ASP ASN THR THR SEQRES 31 C 457 CYS PRO THR ALA VAL LEU GLY ASN VAL ILE ILE SER LEU SEQRES 32 C 457 GLY LYS TYR LEU GLY SER VAL ASN TYR ASN SER GLU GLY SEQRES 33 C 457 ILE ALA ILE GLY PRO PRO VAL PHE THR ASP LYS VAL ASP SEQRES 34 C 457 ILE SER SER GLN ILE SER SER MET ASN GLN SER LEU GLN SEQRES 35 C 457 GLN SER LYS ASP TYR ILE LYS GLU ALA GLN ARG LEU LEU SEQRES 36 C 457 ASP THR SEQRES 1 D 124 VAL GLN LEU VAL GLU SER GLY GLY GLY LEU VAL GLN ALA SEQRES 2 D 124 GLY GLY SER LEU ARG LEU SER CYS ALA ALA SER GLY ARG SEQRES 3 D 124 THR PHE SER SER TYR VAL MET GLY TRP TYR ARG GLN ALA SEQRES 4 D 124 PRO GLY LYS GLU ARG GLU PHE VAL ALA ARG ILE SER TRP SEQRES 5 D 124 SER GLY GLY HIS THR HIS SER ALA ASP SER VAL LYS GLY SEQRES 6 D 124 ARG PHE THR ILE SER ARG ASP ASN ALA LYS ASN THR VAL SEQRES 7 D 124 TYR LEU GLN MET ASN SER LEU LYS PRO ASP ASP THR ALA SEQRES 8 D 124 ILE TYR TYR CYS ASN ALA GLU PRO THR VAL THR GLY SER SEQRES 9 D 124 TRP PHE THR PRO GLY LEU GLN SER TYR TRP GLY GLN GLY SEQRES 10 D 124 THR GLN VAL THR VAL SER SER SEQRES 1 E 124 VAL GLN LEU VAL GLU SER GLY GLY GLY LEU VAL GLN ALA SEQRES 2 E 124 GLY GLY SER LEU ARG LEU SER CYS ALA ALA SER GLY ARG SEQRES 3 E 124 THR PHE SER SER TYR VAL MET GLY TRP TYR ARG GLN ALA SEQRES 4 E 124 PRO GLY LYS GLU ARG GLU PHE VAL ALA ARG ILE SER TRP SEQRES 5 E 124 SER GLY GLY HIS THR HIS SER ALA ASP SER VAL LYS GLY SEQRES 6 E 124 ARG PHE THR ILE SER ARG ASP ASN ALA LYS ASN THR VAL SEQRES 7 E 124 TYR LEU GLN MET ASN SER LEU LYS PRO ASP ASP THR ALA SEQRES 8 E 124 ILE TYR TYR CYS ASN ALA GLU PRO THR VAL THR GLY SER SEQRES 9 E 124 TRP PHE THR PRO GLY LEU GLN SER TYR TRP GLY GLN GLY SEQRES 10 E 124 THR GLN VAL THR VAL SER SER SEQRES 1 F 124 VAL GLN LEU VAL GLU SER GLY GLY GLY LEU VAL GLN ALA SEQRES 2 F 124 GLY GLY SER LEU ARG LEU SER CYS ALA ALA SER GLY ARG SEQRES 3 F 124 THR PHE SER SER TYR VAL MET GLY TRP TYR ARG GLN ALA SEQRES 4 F 124 PRO GLY LYS GLU ARG GLU PHE VAL ALA ARG ILE SER TRP SEQRES 5 F 124 SER GLY GLY HIS THR HIS SER ALA ASP SER VAL LYS GLY SEQRES 6 F 124 ARG PHE THR ILE SER ARG ASP ASN ALA LYS ASN THR VAL SEQRES 7 F 124 TYR LEU GLN MET ASN SER LEU LYS PRO ASP ASP THR ALA SEQRES 8 F 124 ILE TYR TYR CYS ASN ALA GLU PRO THR VAL THR GLY SER SEQRES 9 F 124 TRP PHE THR PRO GLY LEU GLN SER TYR TRP GLY GLN GLY SEQRES 10 F 124 THR GLN VAL THR VAL SER SER HET NAG A 501 14 HET NAG A 502 14 HET NAG A 503 14 HET NAG A 504 14 HET NAG B 501 14 HET NAG B 502 14 HET NAG B 503 14 HET NAG B 504 14 HET NAG C 501 14 HET NAG C 502 14 HET NAG C 503 14 HET NAG C 504 14 HETNAM NAG 2-ACETAMIDO-2-DEOXY-BETA-D-GLUCOPYRANOSE HETSYN NAG N-ACETYL-BETA-D-GLUCOSAMINE; 2-ACETAMIDO-2-DEOXY-BETA- HETSYN 2 NAG D-GLUCOSE; 2-ACETAMIDO-2-DEOXY-D-GLUCOSE; 2-ACETAMIDO- HETSYN 3 NAG 2-DEOXY-GLUCOSE; N-ACETYL-D-GLUCOSAMINE FORMUL 7 NAG 12(C8 H15 N O6) HELIX 1 AA1 HIS A 29 SER A 34 1 6 HELIX 2 AA2 VAL A 65 THR A 72 5 8 HELIX 3 AA3 SER A 74 ASN A 100 1 27 HELIX 4 AA4 GLY A 117 GLY A 121 1 5 HELIX 5 AA5 ALA A 125 ASN A 145 1 21 HELIX 6 AA6 LEU A 147 SER A 153 1 7 HELIX 7 AA7 LEU A 175 ASN A 182 1 8 HELIX 8 AA8 ASN A 182 ILE A 187 1 6 HELIX 9 AA9 SER A 191 GLY A 215 1 25 HELIX 10 AB1 THR A 227 SER A 232 1 6 HELIX 11 AB2 GLN A 233 GLY A 236 5 4 HELIX 12 AB3 ASN A 238 THR A 245 1 8 HELIX 13 AB4 ASP A 252 SER A 259 1 8 HELIX 14 AB5 ILE A 328 CYS A 331 5 4 HELIX 15 AB6 THR A 349 THR A 357 1 9 HELIX 16 AB7 SER A 359 CYS A 363 5 5 HELIX 17 AB8 ASN A 437 GLU A 441 5 5 HELIX 18 AB9 ASP A 452 LEU A 481 1 30 HELIX 19 AC1 HIS B 29 LYS B 35 1 7 HELIX 20 AC2 VAL B 65 THR B 72 5 8 HELIX 21 AC3 SER B 74 ASN B 100 1 27 HELIX 22 AC4 GLY B 117 GLY B 121 1 5 HELIX 23 AC5 ALA B 125 ASN B 145 1 21 HELIX 24 AC6 LEU B 147 GLU B 152 1 6 HELIX 25 AC7 LEU B 175 ASN B 182 1 8 HELIX 26 AC8 ASN B 182 ILE B 187 1 6 HELIX 27 AC9 SER B 191 GLY B 215 1 25 HELIX 28 AD1 THR B 227 SER B 232 1 6 HELIX 29 AD2 GLN B 233 GLY B 236 5 4 HELIX 30 AD3 ASN B 238 THR B 245 1 8 HELIX 31 AD4 ASP B 252 SER B 259 1 8 HELIX 32 AD5 ILE B 328 CYS B 331 5 4 HELIX 33 AD6 THR B 349 THR B 357 1 9 HELIX 34 AD7 SER B 359 CYS B 363 5 5 HELIX 35 AD8 ASN B 437 GLY B 442 1 6 HELIX 36 AD9 ASP B 452 LEU B 481 1 30 HELIX 37 AE1 HIS C 29 LYS C 35 1 7 HELIX 38 AE2 VAL C 65 THR C 72 5 8 HELIX 39 AE3 SER C 74 ASN C 100 1 27 HELIX 40 AE4 GLY C 117 GLY C 121 1 5 HELIX 41 AE5 ALA C 125 ASN C 145 1 21 HELIX 42 AE6 LEU C 147 SER C 153 1 7 HELIX 43 AE7 LEU C 175 ASN C 182 1 8 HELIX 44 AE8 ASN C 182 ILE C 187 1 6 HELIX 45 AE9 SER C 191 GLY C 215 1 25 HELIX 46 AF1 THR C 227 SER C 232 1 6 HELIX 47 AF2 GLN C 233 GLY C 236 5 4 HELIX 48 AF3 ASN C 238 THR C 245 1 8 HELIX 49 AF4 ASP C 252 SER C 259 1 8 HELIX 50 AF5 ILE C 328 CYS C 331 5 4 HELIX 51 AF6 THR C 349 THR C 357 1 9 HELIX 52 AF7 SER C 359 CYS C 363 5 5 HELIX 53 AF8 ASN C 437 GLY C 442 1 6 HELIX 54 AF9 ASP C 452 LEU C 481 1 30 HELIX 55 AG1 GLY D 25 SER D 29 5 5 HELIX 56 AG2 SER D 62 LYS D 64 5 3 HELIX 57 AG3 LYS D 86 THR D 90 5 5 HELIX 58 AG4 GLY E 25 SER E 29 5 5 HELIX 59 AG5 SER E 62 LYS E 64 5 3 HELIX 60 AG6 LYS E 86 THR E 90 5 5 HELIX 61 AG7 GLY F 25 SER F 29 5 5 HELIX 62 AG8 SER F 62 LYS F 64 5 3 HELIX 63 AG9 LYS F 86 THR F 90 5 5 SHEET 1 AA1 7 LEU A 332 ILE A 333 0 SHEET 2 AA1 7 SER A 337 CYS A 340 -1 O ILE A 339 N LEU A 332 SHEET 3 AA1 7 LEU A 38 ILE A 48 1 N LYS A 45 O VAL A 338 SHEET 4 AA1 7 THR A 286 PRO A 298 -1 O LEU A 297 N VAL A 39 SHEET 5 AA1 7 PHE A 315 ARG A 319 -1 O ILE A 316 N GLN A 294 SHEET 6 AA1 7 LEU A 322 ILE A 326 -1 O LEU A 322 N ARG A 319 SHEET 7 AA1 7 ALA A 345 THR A 346 -1 O THR A 346 N ASN A 325 SHEET 1 AA2 5 VAL A 158 GLN A 162 0 SHEET 2 AA2 5 THR A 168 THR A 173 -1 O VAL A 169 N LEU A 161 SHEET 3 AA2 5 LEU A 53 LYS A 60 1 N LYS A 60 O LEU A 172 SHEET 4 AA2 5 TYR A 275 PHE A 282 -1 O VAL A 280 N LYS A 55 SHEET 5 AA2 5 THR A 263 VAL A 269 -1 N ILE A 267 O ILE A 277 SHEET 1 AA3 4 THR A 101 ASP A 103 0 SHEET 2 AA3 4 VAL A 113 ALA A 116 -1 O ILE A 114 N HIS A 102 SHEET 3 AA3 4 VAL C 425 SER C 428 1 O SER C 428 N MET A 115 SHEET 4 AA3 4 THR C 419 LEU C 422 -1 N LEU C 422 O VAL C 425 SHEET 1 AA4 4 ALA A 123 THR A 124 0 SHEET 2 AA4 4 PHE C 376 SER C 379 -1 O LEU C 378 N ALA A 123 SHEET 3 AA4 4 VAL C 382 ALA C 385 -1 O VAL C 382 N SER C 379 SHEET 4 AA4 4 LEU C 410 ILE C 412 -1 O LEU C 410 N ALA C 385 SHEET 1 AA5 7 TYR A 248 ALA A 249 0 SHEET 2 AA5 7 THR D 57 ALA D 60 -1 O SER D 59 N ALA A 249 SHEET 3 AA5 7 GLU D 45 ILE D 50 -1 N ARG D 49 O HIS D 58 SHEET 4 AA5 7 TYR D 31 GLN D 38 -1 N ARG D 37 O GLU D 45 SHEET 5 AA5 7 ALA D 91 PRO D 99 -1 O TYR D 94 N TYR D 36 SHEET 6 AA5 7 THR D 118 SER D 123 -1 O THR D 118 N TYR D 93 SHEET 7 AA5 7 GLY D 9 GLN D 12 1 N VAL D 11 O SER D 123 SHEET 1 AA6 6 TYR A 248 ALA A 249 0 SHEET 2 AA6 6 THR D 57 ALA D 60 -1 O SER D 59 N ALA A 249 SHEET 3 AA6 6 GLU D 45 ILE D 50 -1 N ARG D 49 O HIS D 58 SHEET 4 AA6 6 TYR D 31 GLN D 38 -1 N ARG D 37 O GLU D 45 SHEET 5 AA6 6 ALA D 91 PRO D 99 -1 O TYR D 94 N TYR D 36 SHEET 6 AA6 6 TYR D 113 TRP D 114 -1 O TYR D 113 N ALA D 97 SHEET 1 AA7 3 PHE A 301 ASN A 303 0 SHEET 2 AA7 3 SER A 306 SER A 310 -1 O SER A 306 N ASN A 303 SHEET 3 AA7 3 ARG A 365 LEU A 367 -1 O GLU A 366 N ILE A 309 SHEET 1 AA8 4 LEU A 410 ILE A 412 0 SHEET 2 AA8 4 VAL A 382 ALA A 385 -1 N ALA A 385 O LEU A 410 SHEET 3 AA8 4 PHE A 376 SER A 379 -1 N ALA A 377 O PHE A 384 SHEET 4 AA8 4 ALA B 123 THR B 124 -1 O ALA B 123 N LEU A 378 SHEET 1 AA9 4 THR A 419 LEU A 422 0 SHEET 2 AA9 4 VAL A 425 SER A 428 -1 O VAL A 425 N LEU A 422 SHEET 3 AA9 4 VAL B 113 ALA B 116 1 O MET B 115 N SER A 428 SHEET 4 AA9 4 THR B 101 ASP B 103 -1 N HIS B 102 O ILE B 114 SHEET 1 AB1 7 LEU B 332 ILE B 333 0 SHEET 2 AB1 7 SER B 337 CYS B 340 -1 O ILE B 339 N LEU B 332 SHEET 3 AB1 7 LEU B 38 ILE B 48 1 N LYS B 45 O VAL B 338 SHEET 4 AB1 7 THR B 286 PRO B 298 -1 O LEU B 297 N VAL B 39 SHEET 5 AB1 7 PHE B 315 ARG B 319 -1 O ILE B 316 N GLN B 294 SHEET 6 AB1 7 LEU B 322 ILE B 326 -1 O LEU B 322 N ARG B 319 SHEET 7 AB1 7 ALA B 345 THR B 346 -1 O THR B 346 N ASN B 325 SHEET 1 AB2 5 VAL B 158 GLN B 162 0 SHEET 2 AB2 5 THR B 168 THR B 173 -1 O VAL B 169 N LEU B 161 SHEET 3 AB2 5 LEU B 53 LYS B 60 1 N LYS B 60 O LEU B 172 SHEET 4 AB2 5 TYR B 275 PHE B 282 -1 O VAL B 280 N LYS B 55 SHEET 5 AB2 5 THR B 263 VAL B 269 -1 N THR B 263 O TYR B 281 SHEET 1 AB3 7 TYR B 248 ALA B 249 0 SHEET 2 AB3 7 THR E 57 ALA E 60 -1 O SER E 59 N ALA B 249 SHEET 3 AB3 7 GLU E 45 ILE E 50 -1 N ARG E 49 O HIS E 58 SHEET 4 AB3 7 TYR E 31 GLN E 38 -1 N ARG E 37 O GLU E 45 SHEET 5 AB3 7 ALA E 91 PRO E 99 -1 O TYR E 94 N TYR E 36 SHEET 6 AB3 7 THR E 118 SER E 123 -1 O THR E 118 N TYR E 93 SHEET 7 AB3 7 GLY E 9 GLN E 12 1 N VAL E 11 O SER E 123 SHEET 1 AB4 6 TYR B 248 ALA B 249 0 SHEET 2 AB4 6 THR E 57 ALA E 60 -1 O SER E 59 N ALA B 249 SHEET 3 AB4 6 GLU E 45 ILE E 50 -1 N ARG E 49 O HIS E 58 SHEET 4 AB4 6 TYR E 31 GLN E 38 -1 N ARG E 37 O GLU E 45 SHEET 5 AB4 6 ALA E 91 PRO E 99 -1 O TYR E 94 N TYR E 36 SHEET 6 AB4 6 TYR E 113 TRP E 114 -1 O TYR E 113 N ALA E 97 SHEET 1 AB5 3 PHE B 301 ASN B 303 0 SHEET 2 AB5 3 SER B 306 SER B 310 -1 O TRP B 308 N PHE B 301 SHEET 3 AB5 3 ARG B 365 LEU B 367 -1 O GLU B 366 N ILE B 309 SHEET 1 AB6 4 LEU B 410 ILE B 412 0 SHEET 2 AB6 4 VAL B 382 ALA B 385 -1 N ALA B 385 O LEU B 410 SHEET 3 AB6 4 PHE B 376 SER B 379 -1 N SER B 379 O VAL B 382 SHEET 4 AB6 4 ALA C 123 THR C 124 -1 O ALA C 123 N LEU B 378 SHEET 1 AB7 4 THR B 419 LEU B 422 0 SHEET 2 AB7 4 VAL B 425 SER B 428 -1 O VAL B 425 N LEU B 422 SHEET 3 AB7 4 VAL C 113 ALA C 116 1 O MET C 115 N SER B 428 SHEET 4 AB7 4 THR C 101 ASP C 103 -1 N HIS C 102 O ILE C 114 SHEET 1 AB8 7 LEU C 332 ILE C 333 0 SHEET 2 AB8 7 SER C 337 CYS C 340 -1 O ILE C 339 N LEU C 332 SHEET 3 AB8 7 LEU C 38 ILE C 48 1 N LYS C 45 O VAL C 338 SHEET 4 AB8 7 THR C 286 PRO C 298 -1 O LEU C 297 N VAL C 39 SHEET 5 AB8 7 PHE C 315 ARG C 319 -1 O ILE C 316 N GLN C 294 SHEET 6 AB8 7 LEU C 322 ILE C 326 -1 O LEU C 322 N ARG C 319 SHEET 7 AB8 7 ALA C 345 THR C 346 -1 O THR C 346 N ASN C 325 SHEET 1 AB9 5 VAL C 158 GLN C 162 0 SHEET 2 AB9 5 THR C 168 THR C 173 -1 O VAL C 169 N LEU C 161 SHEET 3 AB9 5 LEU C 53 LYS C 60 1 N LYS C 60 O LEU C 172 SHEET 4 AB9 5 TYR C 275 PHE C 282 -1 O VAL C 280 N LYS C 55 SHEET 5 AB9 5 THR C 263 VAL C 269 -1 N THR C 263 O TYR C 281 SHEET 1 AC1 7 TYR C 248 ALA C 249 0 SHEET 2 AC1 7 THR F 57 ALA F 60 -1 O SER F 59 N ALA C 249 SHEET 3 AC1 7 GLU F 45 ILE F 50 -1 N ARG F 49 O HIS F 58 SHEET 4 AC1 7 TYR F 31 GLN F 38 -1 N ARG F 37 O GLU F 45 SHEET 5 AC1 7 ALA F 91 PRO F 99 -1 O TYR F 94 N TYR F 36 SHEET 6 AC1 7 THR F 118 SER F 123 -1 O THR F 118 N TYR F 93 SHEET 7 AC1 7 GLY F 9 GLN F 12 1 N GLY F 9 O THR F 121 SHEET 1 AC2 6 TYR C 248 ALA C 249 0 SHEET 2 AC2 6 THR F 57 ALA F 60 -1 O SER F 59 N ALA C 249 SHEET 3 AC2 6 GLU F 45 ILE F 50 -1 N ARG F 49 O HIS F 58 SHEET 4 AC2 6 TYR F 31 GLN F 38 -1 N ARG F 37 O GLU F 45 SHEET 5 AC2 6 ALA F 91 PRO F 99 -1 O TYR F 94 N TYR F 36 SHEET 6 AC2 6 TYR F 113 TRP F 114 -1 O TYR F 113 N ALA F 97 SHEET 1 AC3 3 PHE C 301 ASN C 303 0 SHEET 2 AC3 3 SER C 306 SER C 310 -1 O TRP C 308 N PHE C 301 SHEET 3 AC3 3 ARG C 365 LEU C 367 -1 O GLU C 366 N ILE C 309 SHEET 1 AC4 4 VAL D 4 SER D 6 0 SHEET 2 AC4 4 LEU D 17 ALA D 22 -1 O ALA D 22 N VAL D 4 SHEET 3 AC4 4 THR D 77 MET D 82 -1 O LEU D 80 N LEU D 19 SHEET 4 AC4 4 PHE D 67 ASP D 72 -1 N SER D 70 O TYR D 79 SHEET 1 AC5 4 VAL E 4 SER E 6 0 SHEET 2 AC5 4 LEU E 17 ALA E 22 -1 O ALA E 22 N VAL E 4 SHEET 3 AC5 4 THR E 77 MET E 82 -1 O VAL E 78 N CYS E 21 SHEET 4 AC5 4 PHE E 67 ASP E 72 -1 N SER E 70 O TYR E 79 SHEET 1 AC6 4 VAL F 4 SER F 6 0 SHEET 2 AC6 4 LEU F 17 ALA F 22 -1 O ALA F 22 N VAL F 4 SHEET 3 AC6 4 THR F 77 MET F 82 -1 O VAL F 78 N CYS F 21 SHEET 4 AC6 4 PHE F 67 ASP F 72 -1 N SER F 70 O TYR F 79 SSBOND 1 CYS A 71 CYS A 192 1555 1555 2.04 SSBOND 2 CYS A 331 CYS A 340 1555 1555 2.03 SSBOND 3 CYS A 355 CYS A 363 1555 1555 2.03 SSBOND 4 CYS A 387 CYS A 392 1555 1555 2.03 SSBOND 5 CYS A 394 CYS A 417 1555 1555 2.03 SSBOND 6 CYS B 71 CYS B 192 1555 1555 2.04 SSBOND 7 CYS B 331 CYS B 340 1555 1555 2.03 SSBOND 8 CYS B 355 CYS B 363 1555 1555 2.03 SSBOND 9 CYS B 387 CYS B 392 1555 1555 2.03 SSBOND 10 CYS B 394 CYS B 417 1555 1555 2.04 SSBOND 11 CYS C 71 CYS C 192 1555 1555 2.04 SSBOND 12 CYS C 331 CYS C 340 1555 1555 2.03 SSBOND 13 CYS C 355 CYS C 363 1555 1555 2.03 SSBOND 14 CYS C 387 CYS C 392 1555 1555 2.03 SSBOND 15 CYS C 394 CYS C 417 1555 1555 2.03 SSBOND 16 CYS D 21 CYS D 95 1555 1555 2.04 SSBOND 17 CYS E 21 CYS E 95 1555 1555 2.03 SSBOND 18 CYS F 21 CYS F 95 1555 1555 2.04 LINK ND2 ASN A 67 C1 NAG A 504 1555 1555 1.44 LINK ND2 ASN A 99 C1 NAG A 502 1555 1555 1.44 LINK ND2 ASN A 414 C1 NAG A 503 1555 1555 1.44 LINK ND2 ASN A 464 C1 NAG A 501 1555 1555 1.45 LINK ND2 ASN B 67 C1 NAG B 504 1555 1555 1.44 LINK ND2 ASN B 99 C1 NAG B 502 1555 1555 1.44 LINK ND2 ASN B 414 C1 NAG B 501 1555 1555 1.44 LINK ND2 ASN B 464 C1 NAG B 503 1555 1555 1.45 LINK ND2 ASN C 67 C1 NAG C 504 1555 1555 1.44 LINK ND2 ASN C 99 C1 NAG C 502 1555 1555 1.44 LINK ND2 ASN C 414 C1 NAG C 501 1555 1555 1.44 LINK ND2 ASN C 464 C1 NAG C 503 1555 1555 1.45 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