HEADER MEMBRANE PROTEIN 08-SEP-25 9WP9 TITLE CRYO-EM STRUCTURE OF THE D18:1 S1P-BOUND S1PR3 AND GQ COMPLEX COMPND MOL_ID: 1; COMPND 2 MOLECULE: ENGINEERED G-ALPHA-Q; COMPND 3 CHAIN: A; COMPND 4 ENGINEERED: YES; COMPND 5 MOL_ID: 2; COMPND 6 MOLECULE: GUANINE NUCLEOTIDE-BINDING PROTEIN G(I)/G(S)/G(T) SUBUNIT COMPND 7 BETA-1; COMPND 8 CHAIN: B; COMPND 9 SYNONYM: TRANSDUCIN BETA CHAIN 1; COMPND 10 ENGINEERED: YES; COMPND 11 MOL_ID: 3; COMPND 12 MOLECULE: SCFV16; COMPND 13 CHAIN: C; COMPND 14 ENGINEERED: YES; COMPND 15 MOL_ID: 4; COMPND 16 MOLECULE: GUANINE NUCLEOTIDE-BINDING PROTEIN G(I)/G(S)/G(O) SUBUNIT COMPND 17 GAMMA-2; COMPND 18 CHAIN: G; COMPND 19 SYNONYM: G GAMMA-I; COMPND 20 ENGINEERED: YES; COMPND 21 MOL_ID: 5; COMPND 22 MOLECULE: NB35; COMPND 23 CHAIN: N; COMPND 24 ENGINEERED: YES; COMPND 25 MOL_ID: 6; COMPND 26 MOLECULE: SPHINGOSINE 1-PHOSPHATE RECEPTOR 3; COMPND 27 CHAIN: R; COMPND 28 SYNONYM: S1P RECEPTOR 3,S1P3,ENDOTHELIAL DIFFERENTIATION G-PROTEIN COMPND 29 COUPLED RECEPTOR 3,SPHINGOSINE 1-PHOSPHATE RECEPTOR EDG-3,S1P COMPND 30 RECEPTOR EDG-3; COMPND 31 ENGINEERED: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 3 ORGANISM_TAXID: 9606; SOURCE 4 EXPRESSION_SYSTEM: SPODOPTERA FRUGIPERDA; SOURCE 5 EXPRESSION_SYSTEM_TAXID: 7108; SOURCE 6 MOL_ID: 2; SOURCE 7 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 8 ORGANISM_COMMON: HUMAN; SOURCE 9 ORGANISM_TAXID: 9606; SOURCE 10 GENE: GNB1; SOURCE 11 EXPRESSION_SYSTEM: SPODOPTERA FRUGIPERDA; SOURCE 12 EXPRESSION_SYSTEM_TAXID: 7108; SOURCE 13 MOL_ID: 3; SOURCE 14 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 15 ORGANISM_TAXID: 9606; SOURCE 16 EXPRESSION_SYSTEM: SPODOPTERA FRUGIPERDA; SOURCE 17 EXPRESSION_SYSTEM_TAXID: 7108; SOURCE 18 MOL_ID: 4; SOURCE 19 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 20 ORGANISM_COMMON: HUMAN; SOURCE 21 ORGANISM_TAXID: 9606; SOURCE 22 GENE: GNG2; SOURCE 23 EXPRESSION_SYSTEM: SPODOPTERA FRUGIPERDA; SOURCE 24 EXPRESSION_SYSTEM_TAXID: 7108; SOURCE 25 MOL_ID: 5; SOURCE 26 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 27 ORGANISM_TAXID: 9606; SOURCE 28 EXPRESSION_SYSTEM: SPODOPTERA FRUGIPERDA; SOURCE 29 EXPRESSION_SYSTEM_TAXID: 7108; SOURCE 30 MOL_ID: 6; SOURCE 31 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 32 ORGANISM_COMMON: HUMAN; SOURCE 33 ORGANISM_TAXID: 9606; SOURCE 34 GENE: S1PR3, C9ORF108, C9ORF47, EDG3; SOURCE 35 EXPRESSION_SYSTEM: SPODOPTERA FRUGIPERDA; SOURCE 36 EXPRESSION_SYSTEM_TAXID: 7108 KEYWDS GPCR, SBDD, LIPID, MEMBRANE PROTEIN EXPDTA ELECTRON MICROSCOPY AUTHOR D.IM,H.ASADA,S.IWATA,M.YAMAUCHI,M.HAGIWARA REVDAT 1 26-NOV-25 9WP9 0 JRNL AUTH M.YAMAUCHI,D.IM,S.MAEDA,T.IKUTA,M.TOYOMOTO,H.ASADA,Y.SUGITA, JRNL AUTH 2 J.I.KISHIKAWA,T.NODA,T.KATO,A.INOUE,S.IWATA,M.HAGIWARA JRNL TITL STRUCTURAL INSIGHTS INTO THE G-PROTEIN SUBTYPE SELECTIVITY JRNL TITL 2 REVEALED BY HUMAN SPHINGOSINE-1-PHOSPHATE RECEPTOR 3-G Q JRNL TITL 3 COMPLEXES. JRNL REF PROC.NATL.ACAD.SCI.USA V. 122 21122 2025 JRNL REFN ESSN 1091-6490 JRNL PMID 41252158 JRNL DOI 10.1073/PNAS.2507421122 REMARK 2 REMARK 2 RESOLUTION. 3.25 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 SOFTWARE PACKAGES : CRYOSPARC, 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) : 3.250 REMARK 3 NUMBER OF PARTICLES : 366554 REMARK 3 CTF CORRECTION METHOD : NONE REMARK 3 REMARK 3 EM RECONSTRUCTION MAGNIFICATION CALIBRATION: NULL REMARK 3 REMARK 3 OTHER DETAILS: NULL REMARK 4 REMARK 4 9WP9 COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY PDBJ ON 09-SEP-25. REMARK 100 THE DEPOSITION ID IS D_1300063506. 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 : D18:1 S1P-BOUND S1PR3 IN REMARK 245 COMPLEX WITH GQ REMARK 245 SAMPLE CONCENTRATION (MG ML-1) : 10.00 REMARK 245 SAMPLE SUPPORT DETAILS : NULL REMARK 245 SAMPLE VITRIFICATION DETAILS : NULL REMARK 245 SAMPLE BUFFER : NULL REMARK 245 PH : 7.50 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 : TFS 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) : 1800.00 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) : 5000.00 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: HEXAMERIC REMARK 350 SOFTWARE DETERMINED QUATERNARY STRUCTURE: HEXAMERIC REMARK 350 SOFTWARE USED: PISA REMARK 350 APPLY THE FOLLOWING TO CHAINS: A, B, C, G, N, R 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 53 REMARK 465 ARG A 54 REMARK 465 ILE A 55 REMARK 465 LEU A 56 REMARK 465 HIS A 57 REMARK 465 GLY A 58 REMARK 465 GLY A 59 REMARK 465 SER A 60 REMARK 465 GLY A 61 REMARK 465 GLY A 62 REMARK 465 SER A 63 REMARK 465 GLY A 64 REMARK 465 GLY A 65 REMARK 465 THR A 66 REMARK 465 SER A 67 REMARK 465 GLU A 174 REMARK 465 ASP A 175 REMARK 465 ALA A 176 REMARK 465 THR A 177 REMARK 465 PRO A 178 REMARK 465 GLU A 179 REMARK 465 PRO A 180 REMARK 465 GLY A 181 REMARK 465 MET B -17 REMARK 465 HIS B -16 REMARK 465 HIS B -15 REMARK 465 HIS B -14 REMARK 465 HIS B -13 REMARK 465 HIS B -12 REMARK 465 HIS B -11 REMARK 465 LEU B -10 REMARK 465 GLU B -9 REMARK 465 VAL B -8 REMARK 465 LEU B -7 REMARK 465 PHE B -6 REMARK 465 GLN B -5 REMARK 465 GLY B -4 REMARK 465 PRO B -3 REMARK 465 GLY B -2 REMARK 465 SER B -1 REMARK 465 SER B 0 REMARK 465 GLY B 1 REMARK 465 SER B 2 REMARK 465 GLU B 3 REMARK 465 SER C 120 REMARK 465 SER C 121 REMARK 465 GLY C 122 REMARK 465 GLY C 123 REMARK 465 GLY C 124 REMARK 465 GLY C 125 REMARK 465 SER C 126 REMARK 465 GLY C 127 REMARK 465 GLY C 128 REMARK 465 GLY C 129 REMARK 465 GLY C 130 REMARK 465 SER C 131 REMARK 465 GLY C 132 REMARK 465 GLY C 133 REMARK 465 LYS C 248 REMARK 465 ALA C 249 REMARK 465 ALA C 250 REMARK 465 ALA C 251 REMARK 465 HIS C 252 REMARK 465 HIS C 253 REMARK 465 HIS C 254 REMARK 465 HIS C 255 REMARK 465 HIS C 256 REMARK 465 HIS C 257 REMARK 465 HIS C 258 REMARK 465 HIS C 259 REMARK 465 MET G 1 REMARK 465 ALA G 2 REMARK 465 SER G 3 REMARK 465 ASN G 4 REMARK 465 ASN G 5 REMARK 465 GLU G 63 REMARK 465 LYS G 64 REMARK 465 LYS G 65 REMARK 465 PHE G 66 REMARK 465 PHE G 67 REMARK 465 CYS G 68 REMARK 465 ALA G 69 REMARK 465 ILE G 70 REMARK 465 LEU G 71 REMARK 465 MET N 0 REMARK 465 ASP R -26 REMARK 465 TYR R -25 REMARK 465 LYS R -24 REMARK 465 ASP R -23 REMARK 465 ASP R -22 REMARK 465 ASP R -21 REMARK 465 ASP R -20 REMARK 465 LYS R -19 REMARK 465 GLY R -18 REMARK 465 SER R -17 REMARK 465 HIS R -16 REMARK 465 HIS R -15 REMARK 465 HIS R -14 REMARK 465 HIS R -13 REMARK 465 HIS R -12 REMARK 465 HIS R -11 REMARK 465 HIS R -10 REMARK 465 HIS R -9 REMARK 465 GLY R -8 REMARK 465 SER R -7 REMARK 465 LEU R -6 REMARK 465 GLU R -5 REMARK 465 VAL R -4 REMARK 465 LEU R -3 REMARK 465 PHE R -2 REMARK 465 GLN R -1 REMARK 465 GLY R 0 REMARK 465 PRO R 1 REMARK 465 ALA R 2 REMARK 465 THR R 3 REMARK 465 ALA R 4 REMARK 465 LEU R 5 REMARK 465 PRO R 6 REMARK 465 PRO R 7 REMARK 465 ARG R 8 REMARK 465 LEU R 9 REMARK 465 GLN R 10 REMARK 465 PRO R 11 REMARK 465 VAL R 12 REMARK 465 ARG R 13 REMARK 465 GLY R 14 REMARK 465 GLN R 15 REMARK 465 GLU R 16 REMARK 465 LEU R 28 REMARK 465 ALA R 29 REMARK 465 GLY R 30 REMARK 465 ARG R 31 REMARK 465 LEU R 32 REMARK 465 LYS R 33 REMARK 465 GLU R 34 REMARK 465 ALA R 35 REMARK 465 SER R 36 REMARK 465 GLU R 37 REMARK 465 GLY R 38 REMARK 465 SER R 39 REMARK 465 PHE R 70 REMARK 465 HIS R 71 REMARK 465 ASN R 72 REMARK 465 ARG R 73 REMARK 465 LYS R 303 REMARK 465 GLU R 304 REMARK 465 MET R 305 REMARK 465 ARG R 306 REMARK 465 ARG R 307 REMARK 465 ALA R 308 REMARK 465 PHE R 309 REMARK 465 PHE R 310 REMARK 465 ARG R 311 REMARK 465 LEU R 312 REMARK 465 VAL R 313 REMARK 465 CYS R 314 REMARK 465 ASN R 315 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 VAL A 219 CG1 CG2 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 OD1 ASP R 85 OG SER R 291 2.14 REMARK 500 OG SER R 226 ND2 ASN R 234 2.15 REMARK 500 O ILE R 247 OG SER R 250 2.16 REMARK 500 OG SER B 245 OD1 ASP B 247 2.18 REMARK 500 REMARK 500 REMARK: NULL 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 R 185 CA - CB - SG ANGL. DEV. = 6.7 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 ALA A 203 46.56 -85.91 REMARK 500 HIS B 142 -11.06 73.61 REMARK 500 ASP B 205 23.40 -144.74 REMARK 500 VAL C 48 -54.09 -123.47 REMARK 500 ASP C 223 53.64 -90.76 REMARK 500 PRO G 49 41.81 -83.62 REMARK 500 ASN N 77 93.64 -69.32 REMARK 500 ASP N 90 51.24 -93.10 REMARK 500 ALA R 63 -7.05 -56.60 REMARK 500 CYS R 274 70.81 40.38 REMARK 500 REMARK 500 REMARK: NULL REMARK 900 REMARK 900 RELATED ENTRIES REMARK 900 RELATED ID: EMD-66136 RELATED DB: EMDB REMARK 900 CRYO-EM STRUCTURE OF THE D18:1 S1P-BOUND S1PR3 AND GQ COMPLEX DBREF 9WP9 A 5 246 PDB 9WP9 9WP9 5 246 DBREF 9WP9 B 2 340 UNP P62873 GBB1_HUMAN 2 340 DBREF 9WP9 C 1 259 PDB 9WP9 9WP9 1 259 DBREF 9WP9 G 1 71 UNP P59768 GBG2_HUMAN 1 71 DBREF 9WP9 N 0 128 PDB 9WP9 9WP9 0 128 DBREF 9WP9 R 2 315 UNP Q99500 S1PR3_HUMAN 2 315 SEQADV 9WP9 MET B -17 UNP P62873 INITIATING METHIONINE SEQADV 9WP9 HIS B -16 UNP P62873 EXPRESSION TAG SEQADV 9WP9 HIS B -15 UNP P62873 EXPRESSION TAG SEQADV 9WP9 HIS B -14 UNP P62873 EXPRESSION TAG SEQADV 9WP9 HIS B -13 UNP P62873 EXPRESSION TAG SEQADV 9WP9 HIS B -12 UNP P62873 EXPRESSION TAG SEQADV 9WP9 HIS B -11 UNP P62873 EXPRESSION TAG SEQADV 9WP9 LEU B -10 UNP P62873 EXPRESSION TAG SEQADV 9WP9 GLU B -9 UNP P62873 EXPRESSION TAG SEQADV 9WP9 VAL B -8 UNP P62873 EXPRESSION TAG SEQADV 9WP9 LEU B -7 UNP P62873 EXPRESSION TAG SEQADV 9WP9 PHE B -6 UNP P62873 EXPRESSION TAG SEQADV 9WP9 GLN B -5 UNP P62873 EXPRESSION TAG SEQADV 9WP9 GLY B -4 UNP P62873 EXPRESSION TAG SEQADV 9WP9 PRO B -3 UNP P62873 EXPRESSION TAG SEQADV 9WP9 GLY B -2 UNP P62873 EXPRESSION TAG SEQADV 9WP9 SER B -1 UNP P62873 EXPRESSION TAG SEQADV 9WP9 SER B 0 UNP P62873 EXPRESSION TAG SEQADV 9WP9 GLY B 1 UNP P62873 EXPRESSION TAG SEQADV 9WP9 ASP R -26 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 TYR R -25 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 LYS R -24 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 ASP R -23 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 ASP R -22 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 ASP R -21 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 ASP R -20 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 LYS R -19 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 GLY R -18 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 SER R -17 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 HIS R -16 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 HIS R -15 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 HIS R -14 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 HIS R -13 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 HIS R -12 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 HIS R -11 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 HIS R -10 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 HIS R -9 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 GLY R -8 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 SER R -7 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 LEU R -6 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 GLU R -5 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 VAL R -4 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 LEU R -3 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 PHE R -2 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 GLN R -1 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 GLY R 0 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 PRO R 1 UNP Q99500 EXPRESSION TAG SEQADV 9WP9 GLN R 15 UNP Q99500 ASN 15 CONFLICT SEQRES 1 A 242 VAL SER ALA GLU ASP LYS ALA ALA ALA GLU ARG SER LYS SEQRES 2 A 242 MET ILE ASP LYS ASN LEU ARG GLU ASP GLY GLU LYS ALA SEQRES 3 A 242 ARG ARG THR LEU ARG LEU LEU LEU LEU GLY ALA ASP ASN SEQRES 4 A 242 SER GLY LYS SER THR ILE VAL LYS GLN MET ARG ILE LEU SEQRES 5 A 242 HIS GLY GLY SER GLY GLY SER GLY GLY THR SER GLY ILE SEQRES 6 A 242 PHE GLU THR LYS PHE GLN VAL ASP LYS VAL ASN PHE HIS SEQRES 7 A 242 MET PHE ASP VAL GLY GLY GLN ARG ASP GLU ARG ARG LYS SEQRES 8 A 242 TRP ILE GLN CYS PHE ASN ASP VAL THR ALA ILE ILE PHE SEQRES 9 A 242 VAL VAL ASP SER SER ASP TYR ASN ARG LEU GLN GLU ALA SEQRES 10 A 242 LEU ASN ASP PHE LYS SER ILE TRP ASN ASN ARG TRP LEU SEQRES 11 A 242 ARG THR ILE SER VAL ILE LEU PHE LEU ASN LYS GLN ASP SEQRES 12 A 242 LEU LEU ALA GLU LYS VAL LEU ALA GLY LYS SER LYS ILE SEQRES 13 A 242 GLU ASP TYR PHE PRO GLU PHE ALA ARG TYR THR THR PRO SEQRES 14 A 242 GLU ASP ALA THR PRO GLU PRO GLY GLU ASP PRO ARG VAL SEQRES 15 A 242 THR ARG ALA LYS TYR PHE ILE ARG LYS GLU PHE VAL ASP SEQRES 16 A 242 ILE SER THR ALA SER GLY ASP GLY ARG HIS ILE CYS TYR SEQRES 17 A 242 PRO HIS PHE THR CYS ALA VAL ASP THR GLU ASN ALA ARG SEQRES 18 A 242 ARG ILE PHE ASN ASP CYS LYS ASP ILE ILE LEU GLN MET SEQRES 19 A 242 ASN LEU LYS GLU TYR ASN LEU VAL SEQRES 1 B 358 MET HIS HIS HIS HIS HIS HIS LEU GLU VAL LEU PHE GLN SEQRES 2 B 358 GLY PRO GLY SER SER GLY SER GLU LEU ASP GLN LEU ARG SEQRES 3 B 358 GLN GLU ALA GLU GLN LEU LYS ASN GLN ILE ARG ASP ALA SEQRES 4 B 358 ARG LYS ALA CYS ALA ASP ALA THR LEU SER GLN ILE THR SEQRES 5 B 358 ASN ASN ILE ASP PRO VAL GLY ARG ILE GLN MET ARG THR SEQRES 6 B 358 ARG ARG THR LEU ARG GLY HIS LEU ALA LYS ILE TYR ALA SEQRES 7 B 358 MET HIS TRP GLY THR ASP SER ARG LEU LEU VAL SER ALA SEQRES 8 B 358 SER GLN ASP GLY LYS LEU ILE ILE TRP ASP SER TYR THR SEQRES 9 B 358 THR ASN LYS VAL HIS ALA ILE PRO LEU ARG SER SER TRP SEQRES 10 B 358 VAL MET THR CYS ALA TYR ALA PRO SER GLY ASN TYR VAL SEQRES 11 B 358 ALA CYS GLY GLY LEU ASP ASN ILE CYS SER ILE TYR ASN SEQRES 12 B 358 LEU LYS THR ARG GLU GLY ASN VAL ARG VAL SER ARG GLU SEQRES 13 B 358 LEU ALA GLY HIS THR GLY TYR LEU SER CYS CYS ARG PHE SEQRES 14 B 358 LEU ASP ASP ASN GLN ILE VAL THR SER SER GLY ASP THR SEQRES 15 B 358 THR CYS ALA LEU TRP ASP ILE GLU THR GLY GLN GLN THR SEQRES 16 B 358 THR THR PHE THR GLY HIS THR GLY ASP VAL MET SER LEU SEQRES 17 B 358 SER LEU ALA PRO ASP THR ARG LEU PHE VAL SER GLY ALA SEQRES 18 B 358 CYS ASP ALA SER ALA LYS LEU TRP ASP VAL ARG GLU GLY SEQRES 19 B 358 MET CYS ARG GLN THR PHE THR GLY HIS GLU SER ASP ILE SEQRES 20 B 358 ASN ALA ILE CYS PHE PHE PRO ASN GLY ASN ALA PHE ALA SEQRES 21 B 358 THR GLY SER ASP ASP ALA THR CYS ARG LEU PHE ASP LEU SEQRES 22 B 358 ARG ALA ASP GLN GLU LEU MET THR TYR SER HIS ASP ASN SEQRES 23 B 358 ILE ILE CYS GLY ILE THR SER VAL SER PHE SER LYS SER SEQRES 24 B 358 GLY ARG LEU LEU LEU ALA GLY TYR ASP ASP PHE ASN CYS SEQRES 25 B 358 ASN VAL TRP ASP ALA LEU LYS ALA ASP ARG ALA GLY VAL SEQRES 26 B 358 LEU ALA GLY HIS ASP ASN ARG VAL SER CYS LEU GLY VAL SEQRES 27 B 358 THR ASP ASP GLY MET ALA VAL ALA THR GLY SER TRP ASP SEQRES 28 B 358 SER PHE LEU LYS ILE TRP ASN SEQRES 1 C 259 ASP VAL GLN LEU VAL GLU SER GLY GLY GLY LEU VAL GLN SEQRES 2 C 259 PRO GLY GLY SER ARG LYS LEU SER CYS SER ALA SER GLY SEQRES 3 C 259 PHE ALA PHE SER SER PHE GLY MET HIS TRP VAL ARG GLN SEQRES 4 C 259 ALA PRO GLU LYS GLY LEU GLU TRP VAL ALA TYR ILE SER SEQRES 5 C 259 SER GLY SER GLY THR ILE TYR TYR ALA ASP THR VAL LYS SEQRES 6 C 259 GLY ARG PHE THR ILE SER ARG ASP ASP PRO LYS ASN THR SEQRES 7 C 259 LEU PHE LEU GLN MET THR SER LEU ARG SER GLU ASP THR SEQRES 8 C 259 ALA MET TYR TYR CYS VAL ARG SER ILE TYR TYR TYR GLY SEQRES 9 C 259 SER SER PRO PHE ASP PHE TRP GLY GLN GLY THR THR LEU SEQRES 10 C 259 THR VAL SER SER GLY GLY GLY GLY SER GLY GLY GLY GLY SEQRES 11 C 259 SER GLY GLY GLY GLY SER ASP ILE VAL MET THR GLN ALA SEQRES 12 C 259 THR SER SER VAL PRO VAL THR PRO GLY GLU SER VAL SER SEQRES 13 C 259 ILE SER CYS ARG SER SER LYS SER LEU LEU HIS SER ASN SEQRES 14 C 259 GLY ASN THR TYR LEU TYR TRP PHE LEU GLN ARG PRO GLY SEQRES 15 C 259 GLN SER PRO GLN LEU LEU ILE TYR ARG MET SER ASN LEU SEQRES 16 C 259 ALA SER GLY VAL PRO ASP ARG PHE SER GLY SER GLY SER SEQRES 17 C 259 GLY THR ALA PHE THR LEU THR ILE SER ARG LEU GLU ALA SEQRES 18 C 259 GLU ASP VAL GLY VAL TYR TYR CYS MET GLN HIS LEU GLU SEQRES 19 C 259 TYR PRO LEU THR PHE GLY ALA GLY THR LYS LEU GLU LEU SEQRES 20 C 259 LYS ALA ALA ALA HIS HIS HIS HIS HIS HIS HIS HIS SEQRES 1 G 71 MET ALA SER ASN ASN THR ALA SER ILE ALA GLN ALA ARG SEQRES 2 G 71 LYS LEU VAL GLU GLN LEU LYS MET GLU ALA ASN ILE ASP SEQRES 3 G 71 ARG ILE LYS VAL SER LYS ALA ALA ALA ASP LEU MET ALA SEQRES 4 G 71 TYR CYS GLU ALA HIS ALA LYS GLU ASP PRO LEU LEU THR SEQRES 5 G 71 PRO VAL PRO ALA SER GLU ASN PRO PHE ARG GLU LYS LYS SEQRES 6 G 71 PHE PHE CYS ALA ILE LEU SEQRES 1 N 129 MET GLN VAL GLN LEU GLN GLU SER GLY GLY GLY LEU VAL SEQRES 2 N 129 GLN PRO GLY GLY SER LEU ARG LEU SER CYS ALA ALA SER SEQRES 3 N 129 GLY PHE THR PHE SER ASN TYR LYS MET ASN TRP VAL ARG SEQRES 4 N 129 GLN ALA PRO GLY LYS GLY LEU GLU TRP VAL SER ASP ILE SEQRES 5 N 129 SER GLN SER GLY ALA SER ILE SER TYR THR GLY SER VAL SEQRES 6 N 129 LYS GLY ARG PHE THR ILE SER ARG ASP ASN ALA LYS ASN SEQRES 7 N 129 THR LEU TYR LEU GLN MET ASN SER LEU LYS PRO GLU ASP SEQRES 8 N 129 THR ALA VAL TYR TYR CYS ALA ARG CYS PRO ALA PRO PHE SEQRES 9 N 129 THR ARG ASP CYS PHE ASP VAL THR SER THR THR TYR ALA SEQRES 10 N 129 TYR ARG GLY GLN GLY THR GLN VAL THR VAL SER SER SEQRES 1 R 342 ASP TYR LYS ASP ASP ASP ASP LYS GLY SER HIS HIS HIS SEQRES 2 R 342 HIS HIS HIS HIS HIS GLY SER LEU GLU VAL LEU PHE GLN SEQRES 3 R 342 GLY PRO ALA THR ALA LEU PRO PRO ARG LEU GLN PRO VAL SEQRES 4 R 342 ARG GLY GLN GLU THR LEU ARG GLU HIS TYR GLN TYR VAL SEQRES 5 R 342 GLY LYS LEU ALA GLY ARG LEU LYS GLU ALA SER GLU GLY SEQRES 6 R 342 SER THR LEU THR THR VAL LEU PHE LEU VAL ILE CYS SER SEQRES 7 R 342 PHE ILE VAL LEU GLU ASN LEU MET VAL LEU ILE ALA ILE SEQRES 8 R 342 TRP LYS ASN ASN LYS PHE HIS ASN ARG MET TYR PHE PHE SEQRES 9 R 342 ILE GLY ASN LEU ALA LEU CYS ASP LEU LEU ALA GLY ILE SEQRES 10 R 342 ALA TYR LYS VAL ASN ILE LEU MET SER GLY LYS LYS THR SEQRES 11 R 342 PHE SER LEU SER PRO THR VAL TRP PHE LEU ARG GLU GLY SEQRES 12 R 342 SER MET PHE VAL ALA LEU GLY ALA SER THR CYS SER LEU SEQRES 13 R 342 LEU ALA ILE ALA ILE GLU ARG HIS LEU THR MET ILE LYS SEQRES 14 R 342 MET ARG PRO TYR ASP ALA ASN LYS ARG HIS ARG VAL PHE SEQRES 15 R 342 LEU LEU ILE GLY MET CYS TRP LEU ILE ALA PHE THR LEU SEQRES 16 R 342 GLY ALA LEU PRO ILE LEU GLY TRP ASN CYS LEU HIS ASN SEQRES 17 R 342 LEU PRO ASP CYS SER THR ILE LEU PRO LEU TYR SER LYS SEQRES 18 R 342 LYS TYR ILE ALA PHE CYS ILE SER ILE PHE THR ALA ILE SEQRES 19 R 342 LEU VAL THR ILE VAL ILE LEU TYR ALA ARG ILE TYR PHE SEQRES 20 R 342 LEU VAL LYS SER SER SER ARG LYS VAL ALA ASN HIS ASN SEQRES 21 R 342 ASN SER GLU ARG SER MET ALA LEU LEU ARG THR VAL VAL SEQRES 22 R 342 ILE VAL VAL SER VAL PHE ILE ALA CYS TRP SER PRO LEU SEQRES 23 R 342 PHE ILE LEU PHE LEU ILE ASP VAL ALA CYS ARG VAL GLN SEQRES 24 R 342 ALA CYS PRO ILE LEU PHE LYS ALA GLN TRP PHE ILE VAL SEQRES 25 R 342 LEU ALA VAL LEU ASN SER ALA MET ASN PRO VAL ILE TYR SEQRES 26 R 342 THR LEU ALA SER LYS GLU MET ARG ARG ALA PHE PHE ARG SEQRES 27 R 342 LEU VAL CYS ASN HET S1P R1001 25 HETNAM S1P (2S,3R,4E)-2-AMINO-3-HYDROXYOCTADEC-4-EN-1-YL HETNAM 2 S1P DIHYDROGEN PHOSPHATE HETSYN S1P SPHINGOSINE 1-PHOSPHATE FORMUL 7 S1P C18 H38 N O5 P HELIX 1 AA1 SER A 6 ARG A 31 1 26 HELIX 2 AA2 ASP A 42 GLY A 45 5 4 HELIX 3 AA3 ASP A 114 ASN A 116 5 3 HELIX 4 AA4 ARG A 117 ASN A 130 1 14 HELIX 5 AA5 LYS A 145 ALA A 155 1 11 HELIX 6 AA6 LYS A 159 PHE A 164 1 6 HELIX 7 AA7 PRO A 165 ALA A 168 5 4 HELIX 8 AA8 ASP A 183 ALA A 203 1 21 HELIX 9 AA9 GLU A 222 TYR A 243 1 22 HELIX 10 AB1 ASP B 5 ALA B 24 1 20 HELIX 11 AB2 THR B 29 THR B 34 1 6 HELIX 12 AB3 ALA C 28 PHE C 32 5 5 HELIX 13 AB4 ALA G 7 ASN G 24 1 18 HELIX 14 AB5 LYS G 29 HIS G 44 1 16 HELIX 15 AB6 ALA G 45 ASP G 48 5 4 HELIX 16 AB7 THR N 28 TYR N 32 5 5 HELIX 17 AB8 LEU R 18 GLY R 26 1 9 HELIX 18 AB9 THR R 42 ALA R 63 1 22 HELIX 19 AC1 ILE R 64 LYS R 66 5 3 HELIX 20 AC2 TYR R 75 THR R 103 1 29 HELIX 21 AC3 SER R 107 LYS R 142 1 36 HELIX 22 AC4 LYS R 150 GLY R 175 1 26 HELIX 23 AC5 SER R 193 ALA R 230 1 38 HELIX 24 AC6 LEU R 241 CYS R 269 1 29 HELIX 25 AC7 LYS R 279 ASN R 294 1 16 SHEET 1 AA1 5 ILE A 69 VAL A 76 0 SHEET 2 AA1 5 VAL A 79 VAL A 86 -1 O PHE A 81 N PHE A 74 SHEET 3 AA1 5 THR A 33 GLY A 40 1 N LEU A 36 O HIS A 82 SHEET 4 AA1 5 ALA A 105 ASP A 111 1 O VAL A 109 N LEU A 39 SHEET 5 AA1 5 SER A 138 ASN A 144 1 O ILE A 140 N PHE A 108 SHEET 1 AA2 4 THR B 47 LEU B 51 0 SHEET 2 AA2 4 LEU B 336 TRP B 339 -1 O LEU B 336 N LEU B 51 SHEET 3 AA2 4 VAL B 327 SER B 331 -1 N THR B 329 O LYS B 337 SHEET 4 AA2 4 VAL B 315 VAL B 320 -1 N CYS B 317 O GLY B 330 SHEET 1 AA3 4 ILE B 58 HIS B 62 0 SHEET 2 AA3 4 LEU B 69 SER B 74 -1 O VAL B 71 N HIS B 62 SHEET 3 AA3 4 LYS B 78 ASP B 83 -1 O ILE B 80 N SER B 72 SHEET 4 AA3 4 ASN B 88 PRO B 94 -1 O ASN B 88 N ASP B 83 SHEET 1 AA4 4 VAL B 100 TYR B 105 0 SHEET 2 AA4 4 TYR B 111 GLY B 116 -1 O ALA B 113 N ALA B 104 SHEET 3 AA4 4 ILE B 120 ASN B 125 -1 O TYR B 124 N VAL B 112 SHEET 4 AA4 4 ARG B 134 ALA B 140 -1 O ARG B 137 N ILE B 123 SHEET 1 AA5 4 LEU B 146 PHE B 151 0 SHEET 2 AA5 4 GLN B 156 SER B 161 -1 O SER B 160 N SER B 147 SHEET 3 AA5 4 CYS B 166 ASP B 170 -1 O ALA B 167 N THR B 159 SHEET 4 AA5 4 GLN B 176 PHE B 180 -1 O THR B 178 N LEU B 168 SHEET 1 AA6 4 SER B 191 LEU B 192 0 SHEET 2 AA6 4 LEU B 198 GLY B 202 -1 O VAL B 200 N SER B 191 SHEET 3 AA6 4 ALA B 208 ASP B 212 -1 O LYS B 209 N SER B 201 SHEET 4 AA6 4 MET B 217 PHE B 222 -1 O ARG B 219 N LEU B 210 SHEET 1 AA7 4 ILE B 229 PHE B 234 0 SHEET 2 AA7 4 ALA B 240 SER B 245 -1 O GLY B 244 N ALA B 231 SHEET 3 AA7 4 CYS B 250 ASP B 254 -1 O PHE B 253 N PHE B 241 SHEET 4 AA7 4 GLN B 259 TYR B 264 -1 O LEU B 261 N LEU B 252 SHEET 1 AA8 4 ILE B 273 SER B 277 0 SHEET 2 AA8 4 LEU B 284 TYR B 289 -1 O GLY B 288 N THR B 274 SHEET 3 AA8 4 CYS B 294 ASP B 298 -1 O TRP B 297 N LEU B 285 SHEET 4 AA8 4 ARG B 304 LEU B 308 -1 O GLY B 306 N VAL B 296 SHEET 1 AA9 4 GLN C 3 SER C 7 0 SHEET 2 AA9 4 SER C 17 SER C 25 -1 O SER C 21 N SER C 7 SHEET 3 AA9 4 THR C 78 THR C 84 -1 O MET C 83 N ARG C 18 SHEET 4 AA9 4 PHE C 68 ASP C 73 -1 N THR C 69 O GLN C 82 SHEET 1 AB1 5 ILE C 58 TYR C 60 0 SHEET 2 AB1 5 LEU C 45 ILE C 51 -1 N TYR C 50 O TYR C 59 SHEET 3 AB1 5 GLY C 33 GLN C 39 -1 N MET C 34 O ILE C 51 SHEET 4 AB1 5 MET C 93 SER C 99 -1 O TYR C 95 N VAL C 37 SHEET 5 AB1 5 THR C 115 THR C 116 -1 O THR C 115 N TYR C 94 SHEET 1 AB2 4 MET C 140 THR C 141 0 SHEET 2 AB2 4 VAL C 155 SER C 161 -1 O ARG C 160 N THR C 141 SHEET 3 AB2 4 ALA C 211 ILE C 216 -1 O ILE C 216 N VAL C 155 SHEET 4 AB2 4 PHE C 203 GLY C 207 -1 N SER C 206 O THR C 213 SHEET 1 AB3 5 SER C 146 PRO C 148 0 SHEET 2 AB3 5 THR C 243 GLU C 246 1 O LYS C 244 N VAL C 147 SHEET 3 AB3 5 GLY C 225 GLN C 231 -1 N GLY C 225 O LEU C 245 SHEET 4 AB3 5 LEU C 174 GLN C 179 -1 N TYR C 175 O MET C 230 SHEET 5 AB3 5 GLN C 186 ILE C 189 -1 O GLN C 186 N LEU C 178 SHEET 1 AB4 4 LEU N 4 SER N 7 0 SHEET 2 AB4 4 LEU N 18 ALA N 24 -1 O ALA N 23 N GLN N 5 SHEET 3 AB4 4 THR N 78 MET N 83 -1 O LEU N 81 N LEU N 20 SHEET 4 AB4 4 PHE N 68 ASP N 73 -1 N SER N 71 O TYR N 80 SHEET 1 AB5 5 LEU N 11 VAL N 12 0 SHEET 2 AB5 5 THR N 122 VAL N 126 1 O THR N 125 N VAL N 12 SHEET 3 AB5 5 ALA N 92 ARG N 98 -1 N TYR N 94 O THR N 122 SHEET 4 AB5 5 MET N 34 GLN N 39 -1 N ASN N 35 O ALA N 97 SHEET 5 AB5 5 LEU N 45 SER N 49 -1 O GLU N 46 N ARG N 38 SSBOND 1 CYS C 22 CYS C 96 1555 1555 2.03 SSBOND 2 CYS C 159 CYS C 229 1555 1555 2.03 SSBOND 3 CYS N 22 CYS N 96 1555 1555 2.04 SSBOND 4 CYS R 178 CYS R 185 1555 1555 2.03 SSBOND 5 CYS R 269 CYS R 274 1555 1555 2.03 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