HEADER MEMBRANE PROTEIN 23-APR-24 8Z9P TITLE CRYO-EM STRUCTURE OF HUMAN GPR4-GI COMPLEX COMPND MOL_ID: 1; COMPND 2 MOLECULE: GUANINE NUCLEOTIDE-BINDING PROTEIN G(I)/G(S)/G(T) SUBUNIT COMPND 3 BETA-1; COMPND 4 CHAIN: B; COMPND 5 SYNONYM: TRANSDUCIN BETA CHAIN 1; COMPND 6 ENGINEERED: YES; COMPND 7 MOL_ID: 2; COMPND 8 MOLECULE: GUANINE NUCLEOTIDE-BINDING PROTEIN G(I)/G(S)/G(O) SUBUNIT COMPND 9 GAMMA-2; COMPND 10 CHAIN: G; COMPND 11 SYNONYM: G GAMMA-I; COMPND 12 ENGINEERED: YES; COMPND 13 MOL_ID: 3; COMPND 14 MOLECULE: G-PROTEIN COUPLED RECEPTOR 4; COMPND 15 CHAIN: R; COMPND 16 SYNONYM: G-PROTEIN COUPLED RECEPTOR 6C.L,GPR6C.L; COMPND 17 ENGINEERED: YES; COMPND 18 MOL_ID: 4; COMPND 19 MOLECULE: GUANINE NUCLEOTIDE-BINDING PROTEIN G(I) SUBUNIT ALPHA-1; COMPND 20 CHAIN: C; COMPND 21 SYNONYM: ADENYLATE CYCLASE-INHIBITING G ALPHA PROTEIN; COMPND 22 ENGINEERED: YES; COMPND 23 MUTATION: YES; COMPND 24 MOL_ID: 5; COMPND 25 MOLECULE: SINGLE FAB CHAIN (SVFV16); COMPND 26 CHAIN: S; COMPND 27 ENGINEERED: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: RATTUS NORVEGICUS; SOURCE 3 ORGANISM_COMMON: NORWAY RAT; SOURCE 4 ORGANISM_TAXID: 10116; SOURCE 5 GENE: GNB1; SOURCE 6 EXPRESSION_SYSTEM: SPODOPTERA FRUGIPERDA; SOURCE 7 EXPRESSION_SYSTEM_TAXID: 7108; SOURCE 8 MOL_ID: 2; SOURCE 9 ORGANISM_SCIENTIFIC: BOS TAURUS; SOURCE 10 ORGANISM_COMMON: DOMESTIC CATTLE; SOURCE 11 ORGANISM_TAXID: 9913; SOURCE 12 GENE: GNG2; SOURCE 13 EXPRESSION_SYSTEM: SPODOPTERA FRUGIPERDA; SOURCE 14 EXPRESSION_SYSTEM_TAXID: 7108; SOURCE 15 MOL_ID: 3; SOURCE 16 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 17 ORGANISM_COMMON: HUMAN; SOURCE 18 ORGANISM_TAXID: 9606; SOURCE 19 GENE: GPR4; SOURCE 20 EXPRESSION_SYSTEM: HOMO SAPIENS; SOURCE 21 EXPRESSION_SYSTEM_COMMON: HUMAN; SOURCE 22 EXPRESSION_SYSTEM_TAXID: 9606; SOURCE 23 MOL_ID: 4; SOURCE 24 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 25 ORGANISM_COMMON: HUMAN; SOURCE 26 ORGANISM_TAXID: 9606; SOURCE 27 GENE: GNAI1; SOURCE 28 EXPRESSION_SYSTEM: SPODOPTERA FRUGIPERDA; SOURCE 29 EXPRESSION_SYSTEM_TAXID: 7108; SOURCE 30 MOL_ID: 5; SOURCE 31 ORGANISM_SCIENTIFIC: SYNTHETIC CONSTRUCT; SOURCE 32 ORGANISM_TAXID: 32630; SOURCE 33 EXPRESSION_SYSTEM: TRICHOPLUSIA NI; SOURCE 34 EXPRESSION_SYSTEM_TAXID: 7111; SOURCE 35 EXPRESSION_SYSTEM_CELL: HI5 KEYWDS GPCR, CLASS A, GPR4-GI, CRYO-EM, PROTEIN SENSING, ACTIVE STATE, KEYWDS 2 MEMBRANE PROTEIN EXPDTA ELECTRON MICROSCOPY AUTHOR L.N.CHEN,H.ZHOU,K.XI REVDAT 1 09-JUL-25 8Z9P 0 JRNL AUTH L.N.CHEN,H.ZHOU,K.XI JRNL TITL CRYO-EM STRUCTURE OF HUMAN GPR4-GI COMPLEX JRNL REF TO BE PUBLISHED JRNL REFN REMARK 2 REMARK 2 RESOLUTION. 2.50 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 SOFTWARE PACKAGES : RELION, 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) : 2.500 REMARK 3 NUMBER OF PARTICLES : 380507 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 8Z9P COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY PDBC ON 25-APR-24. REMARK 100 THE DEPOSITION ID IS D_1300045822. 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 : CRYO-EM STRUCTURE OF HUMAN GPR4 REMARK 245 -GI COMPLEX REMARK 245 SAMPLE CONCENTRATION (MG ML-1) : NULL REMARK 245 SAMPLE SUPPORT DETAILS : NULL REMARK 245 SAMPLE VITRIFICATION DETAILS : NULL REMARK 245 SAMPLE BUFFER : NULL REMARK 245 PH : 6.00 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 : FEI FALCON IV (4K X 4K) REMARK 245 MINIMUM DEFOCUS (NM) : 700.00 REMARK 245 MAXIMUM DEFOCUS (NM) : 1500.00 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) : 5200.00 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: PENTAMERIC REMARK 350 APPLY THE FOLLOWING TO CHAINS: B, G, R, C, S 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 B -4 REMARK 465 GLY B -3 REMARK 465 SER B -2 REMARK 465 LEU B -1 REMARK 465 LEU B 0 REMARK 465 GLN B 1 REMARK 465 ALA G 2 REMARK 465 SER G 3 REMARK 465 ASN G 4 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 C 1 REMARK 465 GLY C 2 REMARK 465 ILE C 55 REMARK 465 ILE C 56 REMARK 465 HIS C 57 REMARK 465 GLU C 58 REMARK 465 ALA C 59 REMARK 465 GLY C 60 REMARK 465 TYR C 61 REMARK 465 SER C 62 REMARK 465 GLU C 63 REMARK 465 GLU C 64 REMARK 465 GLU C 65 REMARK 465 CYS C 66 REMARK 465 LYS C 67 REMARK 465 GLN C 68 REMARK 465 TYR C 69 REMARK 465 LYS C 70 REMARK 465 ALA C 71 REMARK 465 VAL C 72 REMARK 465 VAL C 73 REMARK 465 TYR C 74 REMARK 465 SER C 75 REMARK 465 ASN C 76 REMARK 465 THR C 77 REMARK 465 ILE C 78 REMARK 465 GLN C 79 REMARK 465 SER C 80 REMARK 465 ILE C 81 REMARK 465 ILE C 82 REMARK 465 ALA C 83 REMARK 465 ILE C 84 REMARK 465 ILE C 85 REMARK 465 ARG C 86 REMARK 465 ALA C 87 REMARK 465 MET C 88 REMARK 465 GLY C 89 REMARK 465 ARG C 90 REMARK 465 LEU C 91 REMARK 465 LYS C 92 REMARK 465 ILE C 93 REMARK 465 ASP C 94 REMARK 465 PHE C 95 REMARK 465 GLY C 96 REMARK 465 ASP C 97 REMARK 465 SER C 98 REMARK 465 ALA C 99 REMARK 465 ARG C 100 REMARK 465 ALA C 101 REMARK 465 ASP C 102 REMARK 465 ASP C 103 REMARK 465 ALA C 104 REMARK 465 ARG C 105 REMARK 465 GLN C 106 REMARK 465 LEU C 107 REMARK 465 PHE C 108 REMARK 465 VAL C 109 REMARK 465 LEU C 110 REMARK 465 ALA C 111 REMARK 465 GLY C 112 REMARK 465 ALA C 113 REMARK 465 ALA C 114 REMARK 465 GLU C 115 REMARK 465 GLU C 116 REMARK 465 GLY C 117 REMARK 465 PHE C 118 REMARK 465 MET C 119 REMARK 465 THR C 120 REMARK 465 ALA C 121 REMARK 465 GLU C 122 REMARK 465 LEU C 123 REMARK 465 ALA C 124 REMARK 465 GLY C 125 REMARK 465 VAL C 126 REMARK 465 ILE C 127 REMARK 465 LYS C 128 REMARK 465 ARG C 129 REMARK 465 LEU C 130 REMARK 465 TRP C 131 REMARK 465 LYS C 132 REMARK 465 ASP C 133 REMARK 465 SER C 134 REMARK 465 GLY C 135 REMARK 465 VAL C 136 REMARK 465 GLN C 137 REMARK 465 ALA C 138 REMARK 465 CYS C 139 REMARK 465 PHE C 140 REMARK 465 ASN C 141 REMARK 465 ARG C 142 REMARK 465 SER C 143 REMARK 465 ARG C 144 REMARK 465 GLU C 145 REMARK 465 TYR C 146 REMARK 465 GLN C 147 REMARK 465 LEU C 148 REMARK 465 ASN C 149 REMARK 465 ASP C 150 REMARK 465 SER C 151 REMARK 465 ALA C 152 REMARK 465 ALA C 153 REMARK 465 TYR C 154 REMARK 465 TYR C 155 REMARK 465 LEU C 156 REMARK 465 ASN C 157 REMARK 465 ASP C 158 REMARK 465 LEU C 159 REMARK 465 ASP C 160 REMARK 465 ARG C 161 REMARK 465 ILE C 162 REMARK 465 ALA C 163 REMARK 465 GLN C 164 REMARK 465 PRO C 165 REMARK 465 ASN C 166 REMARK 465 TYR C 167 REMARK 465 ILE C 168 REMARK 465 PRO C 169 REMARK 465 THR C 170 REMARK 465 GLN C 171 REMARK 465 GLN C 172 REMARK 465 ASP C 173 REMARK 465 VAL C 174 REMARK 465 LEU C 175 REMARK 465 ARG C 176 REMARK 465 THR C 177 REMARK 465 ARG C 178 REMARK 465 VAL C 179 REMARK 465 LYS C 180 REMARK 465 THR C 181 REMARK 465 GLY S 122 REMARK 465 GLY S 123 REMARK 465 GLY S 124 REMARK 465 GLY S 125 REMARK 465 SER S 126 REMARK 465 GLY S 127 REMARK 465 GLY S 128 REMARK 465 GLY S 129 REMARK 465 GLY S 130 REMARK 465 SER S 131 REMARK 465 GLY S 132 REMARK 465 GLY S 133 REMARK 465 GLY S 134 REMARK 465 LYS S 248 REMARK 465 GLY S 249 REMARK 465 SER S 250 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 GLN S 113 CG CD OE1 NE2 REMARK 470 LEU S 233 CG CD1 CD2 REMARK 470 GLU S 234 CG CD OE1 OE2 REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: COVALENT BOND LENGTHS 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,2(A3,1X,A1,I4,A1,1X,A4,3X),1X,F6.3) 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 RES CSSEQI ATM2 DEVIATION REMARK 500 CYS B 114 CB CYS B 114 SG -0.115 REMARK 500 CYS B 149 CB CYS B 149 SG -0.156 REMARK 500 CYS B 250 CB CYS B 250 SG -0.097 REMARK 500 GLU B 260 CD GLU B 260 OE2 -0.067 REMARK 500 TYR B 264 CG TYR B 264 CD2 -0.085 REMARK 500 CYS B 271 CB CYS B 271 SG -0.104 REMARK 500 SER B 331 CB SER B 331 OG -0.082 REMARK 500 TYR R 43 CB TYR R 43 CG -0.104 REMARK 500 TYR R 55 CB TYR R 55 CG -0.093 REMARK 500 PHE R 77 CB PHE R 77 CG -0.102 REMARK 500 TYR R 116 CG TYR R 116 CD2 -0.079 REMARK 500 TYR R 116 CE1 TYR R 116 CZ -0.083 REMARK 500 HIS R 121 CB HIS R 121 CG -0.117 REMARK 500 PHE R 169 CB PHE R 169 CG -0.108 REMARK 500 GLU R 261 CG GLU R 261 CD -0.101 REMARK 500 PHE C 189 CB PHE C 189 CG -0.144 REMARK 500 PHE C 199 CB PHE C 199 CG -0.106 REMARK 500 CYS C 214 CB CYS C 214 SG -0.118 REMARK 500 PHE C 215 CB PHE C 215 CG -0.108 REMARK 500 PHE C 354 CG PHE C 354 CD2 0.095 REMARK 500 PHE C 354 CG PHE C 354 CD1 0.113 REMARK 500 ARG S 38 CB ARG S 38 CG -0.167 REMARK 500 TRP S 47 CB TRP S 47 CG -0.114 REMARK 500 TRP S 47 CE2 TRP S 47 CD2 -0.074 REMARK 500 TYR S 59 CG TYR S 59 CD2 -0.083 REMARK 500 CYS S 96 CB CYS S 96 SG -0.193 REMARK 500 CYS S 229 CB CYS S 229 SG -0.106 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 ARG B 22 NE - CZ - NH2 ANGL. DEV. = -3.2 DEGREES REMARK 500 ARG B 49 NE - CZ - NH2 ANGL. DEV. = -4.2 DEGREES REMARK 500 ARG B 197 NE - CZ - NH2 ANGL. DEV. = -3.4 DEGREES REMARK 500 ARG B 219 NE - CZ - NH2 ANGL. DEV. = -3.3 DEGREES REMARK 500 ARG G 27 NE - CZ - NH2 ANGL. DEV. = -4.5 DEGREES REMARK 500 HIS R 85 N - CA - C ANGL. DEV. = -22.0 DEGREES REMARK 500 ARG R 115 NE - CZ - NH1 ANGL. DEV. = 3.4 DEGREES REMARK 500 ARG R 115 NE - CZ - NH2 ANGL. DEV. = -3.3 DEGREES REMARK 500 ARG C 15 NE - CZ - NH1 ANGL. DEV. = -3.1 DEGREES REMARK 500 TYR C 302 CB - CG - CD1 ANGL. DEV. = -4.0 DEGREES REMARK 500 ARG S 18 NE - CZ - NH2 ANGL. DEV. = -4.3 DEGREES REMARK 500 ARG S 67 NE - CZ - NH1 ANGL. DEV. = 4.1 DEGREES REMARK 500 ARG S 67 NE - CZ - NH2 ANGL. DEV. = -3.7 DEGREES REMARK 500 TYR S 175 CB - CG - CD2 ANGL. DEV. = -4.0 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 THR B 164 -0.73 81.13 REMARK 500 THR B 196 -2.19 68.40 REMARK 500 LEU R 52 8.72 -67.01 REMARK 500 HIS R 80 50.84 104.42 REMARK 500 ILE R 84 82.59 -8.09 REMARK 500 HIS R 85 146.98 170.67 REMARK 500 LEU R 123 -4.65 75.13 REMARK 500 ALA R 179 -39.76 -36.04 REMARK 500 VAL R 264 17.62 -140.42 REMARK 500 THR C 4 -53.66 66.93 REMARK 500 TRP C 211 -19.05 -149.63 REMARK 500 THR C 284 -38.06 -34.36 REMARK 500 ILE C 285 -34.71 -38.76 REMARK 500 MET S 192 -55.34 74.36 REMARK 500 ARG S 218 78.09 56.59 REMARK 500 REMARK 500 REMARK: NULL REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: MAIN CHAIN PLANARITY REMARK 500 REMARK 500 THE FOLLOWING RESIDUES HAVE A PSEUDO PLANARITY REMARK 500 TORSION ANGLE, C(I) - CA(I) - N(I+1) - O(I), GREATER REMARK 500 10.0 DEGREES. (M=MODEL NUMBER; RES=RESIDUE NAME; REMARK 500 C=CHAIN IDENTIFIER; SSEQ=SEQUENCE NUMBER; REMARK 500 I=INSERTION CODE). REMARK 500 REMARK 500 M RES CSSEQI ANGLE REMARK 500 LEU B 30 -25.08 REMARK 500 GLU R 175 -10.74 REMARK 500 PRO R 193 -25.04 REMARK 500 ALA R 293 -24.01 REMARK 500 ALA C 226 -27.91 REMARK 500 PHE C 274 -22.63 REMARK 500 PRO S 107 -25.26 REMARK 500 REMARK 500 REMARK: NULL REMARK 900 REMARK 900 RELATED ENTRIES REMARK 900 RELATED ID: EMD-39866 RELATED DB: EMDB REMARK 900 CRYO-EM STRUCTURE OF HUMAN GPR4-GI COMPLEX DBREF 8Z9P B 2 340 UNP P54311 GBB1_RAT 2 340 DBREF 8Z9P G 2 71 UNP P63212 GBG2_BOVIN 2 71 DBREF 8Z9P R 8 310 UNP P46093 GPR4_HUMAN 8 310 DBREF 8Z9P C 1 354 UNP P63096 GNAI1_HUMAN 1 354 DBREF 8Z9P S 1 250 PDB 8Z9P 8Z9P 1 250 SEQADV 8Z9P MET B -4 UNP P54311 INITIATING METHIONINE SEQADV 8Z9P GLY B -3 UNP P54311 EXPRESSION TAG SEQADV 8Z9P SER B -2 UNP P54311 EXPRESSION TAG SEQADV 8Z9P LEU B -1 UNP P54311 EXPRESSION TAG SEQADV 8Z9P LEU B 0 UNP P54311 EXPRESSION TAG SEQADV 8Z9P GLN B 1 UNP P54311 EXPRESSION TAG SEQADV 8Z9P ASN C 47 UNP P63096 SER 47 ENGINEERED MUTATION SEQADV 8Z9P ALA C 203 UNP P63096 GLY 203 ENGINEERED MUTATION SEQADV 8Z9P ALA C 245 UNP P63096 GLU 245 ENGINEERED MUTATION SEQADV 8Z9P SER C 326 UNP P63096 ALA 326 ENGINEERED MUTATION SEQRES 1 B 345 MET GLY SER LEU LEU GLN SER GLU LEU ASP GLN LEU ARG SEQRES 2 B 345 GLN GLU ALA GLU GLN LEU LYS ASN GLN ILE ARG ASP ALA SEQRES 3 B 345 ARG LYS ALA CYS ALA ASP ALA THR LEU SER GLN ILE THR SEQRES 4 B 345 ASN ASN ILE ASP PRO VAL GLY ARG ILE GLN MET ARG THR SEQRES 5 B 345 ARG ARG THR LEU ARG GLY HIS LEU ALA LYS ILE TYR ALA SEQRES 6 B 345 MET HIS TRP GLY THR ASP SER ARG LEU LEU VAL SER ALA SEQRES 7 B 345 SER GLN ASP GLY LYS LEU ILE ILE TRP ASP SER TYR THR SEQRES 8 B 345 THR ASN LYS VAL HIS ALA ILE PRO LEU ARG SER SER TRP SEQRES 9 B 345 VAL MET THR CYS ALA TYR ALA PRO SER GLY ASN TYR VAL SEQRES 10 B 345 ALA CYS GLY GLY LEU ASP ASN ILE CYS SER ILE TYR ASN SEQRES 11 B 345 LEU LYS THR ARG GLU GLY ASN VAL ARG VAL SER ARG GLU SEQRES 12 B 345 LEU ALA GLY HIS THR GLY TYR LEU SER CYS CYS ARG PHE SEQRES 13 B 345 LEU ASP ASP ASN GLN ILE VAL THR SER SER GLY ASP THR SEQRES 14 B 345 THR CYS ALA LEU TRP ASP ILE GLU THR GLY GLN GLN THR SEQRES 15 B 345 THR THR PHE THR GLY HIS THR GLY ASP VAL MET SER LEU SEQRES 16 B 345 SER LEU ALA PRO ASP THR ARG LEU PHE VAL SER GLY ALA SEQRES 17 B 345 CYS ASP ALA SER ALA LYS LEU TRP ASP VAL ARG GLU GLY SEQRES 18 B 345 MET CYS ARG GLN THR PHE THR GLY HIS GLU SER ASP ILE SEQRES 19 B 345 ASN ALA ILE CYS PHE PHE PRO ASN GLY ASN ALA PHE ALA SEQRES 20 B 345 THR GLY SER ASP ASP ALA THR CYS ARG LEU PHE ASP LEU SEQRES 21 B 345 ARG ALA ASP GLN GLU LEU MET THR TYR SER HIS ASP ASN SEQRES 22 B 345 ILE ILE CYS GLY ILE THR SER VAL SER PHE SER LYS SER SEQRES 23 B 345 GLY ARG LEU LEU LEU ALA GLY TYR ASP ASP PHE ASN CYS SEQRES 24 B 345 ASN VAL TRP ASP ALA LEU LYS ALA ASP ARG ALA GLY VAL SEQRES 25 B 345 LEU ALA GLY HIS ASP ASN ARG VAL SER CYS LEU GLY VAL SEQRES 26 B 345 THR ASP ASP GLY MET ALA VAL ALA THR GLY SER TRP ASP SEQRES 27 B 345 SER PHE LEU LYS ILE TRP ASN SEQRES 1 G 70 ALA SER ASN ASN THR ALA SER ILE ALA GLN ALA ARG LYS SEQRES 2 G 70 LEU VAL GLU GLN LEU LYS MET GLU ALA ASN ILE ASP ARG SEQRES 3 G 70 ILE LYS VAL SER LYS ALA ALA ALA ASP LEU MET ALA TYR SEQRES 4 G 70 CYS GLU ALA HIS ALA LYS GLU ASP PRO LEU LEU THR PRO SEQRES 5 G 70 VAL PRO ALA SER GLU ASN PRO PHE ARG GLU LYS LYS PHE SEQRES 6 G 70 PHE CYS ALA ILE LEU SEQRES 1 R 303 GLY CYS HIS VAL ASP SER ARG VAL ASP HIS LEU PHE PRO SEQRES 2 R 303 PRO SER LEU TYR ILE PHE VAL ILE GLY VAL GLY LEU PRO SEQRES 3 R 303 THR ASN CYS LEU ALA LEU TRP ALA ALA TYR ARG GLN VAL SEQRES 4 R 303 GLN GLN ARG ASN GLU LEU GLY VAL TYR LEU MET ASN LEU SEQRES 5 R 303 SER ILE ALA ASP LEU LEU TYR ILE CYS THR LEU PRO LEU SEQRES 6 R 303 TRP VAL ASP TYR PHE LEU HIS HIS ASP ASN TRP ILE HIS SEQRES 7 R 303 GLY PRO GLY SER CYS LYS LEU PHE GLY PHE ILE PHE TYR SEQRES 8 R 303 THR ASN ILE TYR ILE SER ILE ALA PHE LEU CYS CYS ILE SEQRES 9 R 303 SER VAL ASP ARG TYR LEU ALA VAL ALA HIS PRO LEU ARG SEQRES 10 R 303 PHE ALA ARG LEU ARG ARG VAL LYS THR ALA VAL ALA VAL SEQRES 11 R 303 SER SER VAL VAL TRP ALA THR GLU LEU GLY ALA ASN SER SEQRES 12 R 303 ALA PRO LEU PHE HIS ASP GLU LEU PHE ARG ASP ARG TYR SEQRES 13 R 303 ASN HIS THR PHE CYS PHE GLU LYS PHE PRO MET GLU GLY SEQRES 14 R 303 TRP VAL ALA TRP MET ASN LEU TYR ARG VAL PHE VAL GLY SEQRES 15 R 303 PHE LEU PHE PRO TRP ALA LEU MET LEU LEU SER TYR ARG SEQRES 16 R 303 GLY ILE LEU ARG ALA VAL ARG GLY SER VAL SER THR GLU SEQRES 17 R 303 ARG GLN GLU LYS ALA LYS ILE LYS ARG LEU ALA LEU SER SEQRES 18 R 303 LEU ILE ALA ILE VAL LEU VAL CYS PHE ALA PRO TYR HIS SEQRES 19 R 303 VAL LEU LEU LEU SER ARG SER ALA ILE TYR LEU GLY ARG SEQRES 20 R 303 PRO TRP ASP CYS GLY PHE GLU GLU ARG VAL PHE SER ALA SEQRES 21 R 303 TYR HIS SER SER LEU ALA PHE THR SER LEU ASN CYS VAL SEQRES 22 R 303 ALA ASP PRO ILE LEU TYR CYS LEU VAL ASN GLU GLY ALA SEQRES 23 R 303 ARG SER ASP VAL ALA LYS ALA LEU HIS ASN LEU LEU ARG SEQRES 24 R 303 PHE LEU ALA SER SEQRES 1 C 354 MET GLY CYS THR LEU SER ALA GLU ASP LYS ALA ALA VAL SEQRES 2 C 354 GLU ARG SER LYS MET ILE ASP ARG ASN LEU ARG GLU ASP SEQRES 3 C 354 GLY GLU LYS ALA ALA ARG GLU VAL LYS LEU LEU LEU LEU SEQRES 4 C 354 GLY ALA GLY GLU SER GLY LYS ASN THR ILE VAL LYS GLN SEQRES 5 C 354 MET LYS ILE ILE HIS GLU ALA GLY TYR SER GLU GLU GLU SEQRES 6 C 354 CYS LYS GLN TYR LYS ALA VAL VAL TYR SER ASN THR ILE SEQRES 7 C 354 GLN SER ILE ILE ALA ILE ILE ARG ALA MET GLY ARG LEU SEQRES 8 C 354 LYS ILE ASP PHE GLY ASP SER ALA ARG ALA ASP ASP ALA SEQRES 9 C 354 ARG GLN LEU PHE VAL LEU ALA GLY ALA ALA GLU GLU GLY SEQRES 10 C 354 PHE MET THR ALA GLU LEU ALA GLY VAL ILE LYS ARG LEU SEQRES 11 C 354 TRP LYS ASP SER GLY VAL GLN ALA CYS PHE ASN ARG SER SEQRES 12 C 354 ARG GLU TYR GLN LEU ASN ASP SER ALA ALA TYR TYR LEU SEQRES 13 C 354 ASN ASP LEU ASP ARG ILE ALA GLN PRO ASN TYR ILE PRO SEQRES 14 C 354 THR GLN GLN ASP VAL LEU ARG THR ARG VAL LYS THR THR SEQRES 15 C 354 GLY ILE VAL GLU THR HIS PHE THR PHE LYS ASP LEU HIS SEQRES 16 C 354 PHE LYS MET PHE ASP VAL GLY ALA GLN ARG SER GLU ARG SEQRES 17 C 354 LYS LYS TRP ILE HIS CYS PHE GLU GLY VAL THR ALA ILE SEQRES 18 C 354 ILE PHE CYS VAL ALA LEU SER ASP TYR ASP LEU VAL LEU SEQRES 19 C 354 ALA GLU ASP GLU GLU MET ASN ARG MET HIS ALA SER MET SEQRES 20 C 354 LYS LEU PHE ASP SER ILE CYS ASN ASN LYS TRP PHE THR SEQRES 21 C 354 ASP THR SER ILE ILE LEU PHE LEU ASN LYS LYS ASP LEU SEQRES 22 C 354 PHE GLU GLU LYS ILE LYS LYS SER PRO LEU THR ILE CYS SEQRES 23 C 354 TYR PRO GLU TYR ALA GLY SER ASN THR TYR GLU GLU ALA SEQRES 24 C 354 ALA ALA TYR ILE GLN CYS GLN PHE GLU ASP LEU ASN LYS SEQRES 25 C 354 ARG LYS ASP THR LYS GLU ILE TYR THR HIS PHE THR CYS SEQRES 26 C 354 SER THR ASP THR LYS ASN VAL GLN PHE VAL PHE ASP ALA SEQRES 27 C 354 VAL THR ASP VAL ILE ILE LYS ASN ASN LEU LYS ASP CYS SEQRES 28 C 354 GLY LEU PHE SEQRES 1 S 250 ASP VAL GLN LEU VAL GLU SER GLY GLY GLY LEU VAL GLN SEQRES 2 S 250 PRO GLY GLY SER ARG LYS LEU SER CYS SER ALA SER GLY SEQRES 3 S 250 PHE ALA PHE SER SER PHE GLY MET HIS TRP VAL ARG GLN SEQRES 4 S 250 ALA PRO GLU LYS GLY LEU GLU TRP VAL ALA TYR ILE SER SEQRES 5 S 250 SER GLY SER GLY THR ILE TYR TYR ALA ASP THR VAL LYS SEQRES 6 S 250 GLY ARG PHE THR ILE SER ARG ASP ASP PRO LYS ASN THR SEQRES 7 S 250 LEU PHE LEU GLN MET THR SER LEU ARG SER GLU ASP THR SEQRES 8 S 250 ALA MET TYR TYR CYS VAL ARG SER ILE TYR TYR TYR GLY SEQRES 9 S 250 SER SER PRO PHE ASP PHE TRP GLY GLN GLY THR THR LEU SEQRES 10 S 250 THR VAL SER SER GLY GLY GLY GLY SER GLY GLY GLY GLY SEQRES 11 S 250 SER GLY GLY GLY GLY SER ASP ILE VAL MET THR GLN ALA SEQRES 12 S 250 THR SER SER VAL PRO VAL THR PRO GLY GLU SER VAL SER SEQRES 13 S 250 ILE SER CYS ARG SER SER LYS SER LEU LEU HIS SER ASN SEQRES 14 S 250 GLY ASN THR TYR LEU TYR TRP PHE LEU GLN ARG PRO GLY SEQRES 15 S 250 GLN SER PRO GLN LEU LEU ILE TYR ARG MET SER ASN LEU SEQRES 16 S 250 ALA SER GLY VAL PRO ASP ARG PHE SER GLY SER GLY SER SEQRES 17 S 250 GLY THR ALA PHE THR LEU THR ILE SER ARG LEU GLU ALA SEQRES 18 S 250 GLU ASP VAL GLY VAL TYR TYR CYS MET GLN HIS LEU GLU SEQRES 19 S 250 TYR PRO LEU THR PHE GLY ALA GLY THR LYS LEU GLU LEU SEQRES 20 S 250 LYS GLY SER HET CLR R 401 28 HET CLR R 402 28 HETNAM CLR CHOLESTEROL FORMUL 6 CLR 2(C27 H46 O) HELIX 1 AA1 SER B 2 ALA B 24 1 23 HELIX 2 AA2 THR B 29 THR B 34 1 6 HELIX 3 AA3 ASN B 35 ILE B 37 5 3 HELIX 4 AA4 THR G 6 ASN G 24 1 19 HELIX 5 AA5 LYS G 29 HIS G 44 1 16 HELIX 6 AA6 ALA G 45 ASP G 48 5 4 HELIX 7 AA7 PRO G 55 ASN G 59 5 5 HELIX 8 AA8 LEU R 18 GLN R 48 1 31 HELIX 9 AA9 ARG R 49 LEU R 52 5 4 HELIX 10 AB1 GLY R 53 HIS R 79 1 27 HELIX 11 AB2 PRO R 87 HIS R 121 1 35 HELIX 12 AB3 PHE R 125 ARG R 129 5 5 HELIX 13 AB4 ARG R 130 ASN R 149 1 20 HELIX 14 AB5 SER R 150 HIS R 155 5 6 HELIX 15 AB6 GLU R 175 VAL R 188 1 14 HELIX 16 AB7 PHE R 190 GLY R 210 1 21 HELIX 17 AB8 GLU R 215 ARG R 254 1 40 HELIX 18 AB9 CYS R 258 ARG R 263 1 6 HELIX 19 AC1 VAL R 264 THR R 275 1 12 HELIX 20 AC2 LEU R 277 CYS R 287 1 11 HELIX 21 AC3 ASN R 290 SER R 310 1 21 HELIX 22 AC4 SER C 6 GLU C 33 1 28 HELIX 23 AC5 GLY C 45 MET C 53 1 9 HELIX 24 AC6 GLU C 207 GLU C 216 5 10 HELIX 25 AC7 ALA C 226 ASP C 231 5 6 HELIX 26 AC8 ARG C 242 ASN C 255 1 14 HELIX 27 AC9 LYS C 270 ILE C 278 1 9 HELIX 28 AD1 PRO C 282 CYS C 286 5 5 HELIX 29 AD2 THR C 295 ASP C 309 1 15 HELIX 30 AD3 LYS C 330 CYS C 351 1 22 HELIX 31 AD4 ALA S 28 PHE S 32 5 5 HELIX 32 AD5 SER S 53 GLY S 56 5 4 HELIX 33 AD6 ASP S 74 LYS S 76 5 3 HELIX 34 AD7 ARG S 87 THR S 91 5 5 HELIX 35 AD8 GLU S 220 VAL S 224 5 5 SHEET 1 AA1 4 ARG B 46 LEU B 51 0 SHEET 2 AA1 4 LEU B 336 ASN B 340 -1 O ILE B 338 N ARG B 48 SHEET 3 AA1 4 VAL B 327 SER B 331 -1 N VAL B 327 O TRP B 339 SHEET 4 AA1 4 VAL B 315 VAL B 320 -1 N GLY B 319 O ALA B 328 SHEET 1 AA2 4 ILE B 58 TRP B 63 0 SHEET 2 AA2 4 LEU B 69 SER B 74 -1 O ALA B 73 N ALA B 60 SHEET 3 AA2 4 LYS B 78 ASP B 83 -1 O TRP B 82 N LEU B 70 SHEET 4 AA2 4 ASN B 88 PRO B 94 -1 O VAL B 90 N ILE B 81 SHEET 1 AA3 4 VAL B 100 TYR B 105 0 SHEET 2 AA3 4 TYR B 111 GLY B 116 -1 O GLY B 115 N MET B 101 SHEET 3 AA3 4 ILE B 120 ASN B 125 -1 O TYR B 124 N VAL B 112 SHEET 4 AA3 4 ARG B 134 ALA B 140 -1 O SER B 136 N ILE B 123 SHEET 1 AA4 4 LEU B 146 ASP B 153 0 SHEET 2 AA4 4 GLN B 156 SER B 161 -1 O VAL B 158 N ARG B 150 SHEET 3 AA4 4 CYS B 166 ASP B 170 -1 O ALA B 167 N THR B 159 SHEET 4 AA4 4 GLN B 176 PHE B 180 -1 O THR B 177 N LEU B 168 SHEET 1 AA5 4 VAL B 187 LEU B 192 0 SHEET 2 AA5 4 LEU B 198 ALA B 203 -1 O GLY B 202 N MET B 188 SHEET 3 AA5 4 SER B 207 ASP B 212 -1 O TRP B 211 N PHE B 199 SHEET 4 AA5 4 MET B 217 THR B 223 -1 O ARG B 219 N LEU B 210 SHEET 1 AA6 4 ILE B 229 PHE B 234 0 SHEET 2 AA6 4 ALA B 240 SER B 245 -1 O GLY B 244 N ALA B 231 SHEET 3 AA6 4 CYS B 250 ASP B 254 -1 O PHE B 253 N PHE B 241 SHEET 4 AA6 4 GLN B 259 TYR B 264 -1 O LEU B 261 N LEU B 252 SHEET 1 AA7 4 ILE B 273 PHE B 278 0 SHEET 2 AA7 4 LEU B 284 TYR B 289 -1 O LEU B 286 N SER B 277 SHEET 3 AA7 4 CYS B 294 ASP B 298 -1 O TRP B 297 N LEU B 285 SHEET 4 AA7 4 ARG B 304 LEU B 308 -1 O LEU B 308 N CYS B 294 SHEET 1 AA8 2 LEU R 158 ARG R 160 0 SHEET 2 AA8 2 THR R 166 CYS R 168 -1 O PHE R 167 N PHE R 159 SHEET 1 AA9 6 VAL C 185 PHE C 191 0 SHEET 2 AA9 6 LEU C 194 ASP C 200 -1 O PHE C 196 N PHE C 189 SHEET 3 AA9 6 VAL C 34 GLY C 40 1 N LEU C 36 O PHE C 199 SHEET 4 AA9 6 ALA C 220 VAL C 225 1 O ILE C 222 N LEU C 37 SHEET 5 AA9 6 SER C 263 LEU C 268 1 O ILE C 265 N ILE C 221 SHEET 6 AA9 6 ILE C 319 PHE C 323 1 O TYR C 320 N LEU C 266 SHEET 1 AB1 2 VAL C 233 LEU C 234 0 SHEET 2 AB1 2 ASP C 237 ASN C 241 -1 O MET C 240 N LEU C 234 SHEET 1 AB2 4 GLN S 3 SER S 7 0 SHEET 2 AB2 4 ARG S 18 SER S 25 -1 O SER S 25 N GLN S 3 SHEET 3 AB2 4 THR S 78 MET S 83 -1 O MET S 83 N ARG S 18 SHEET 4 AB2 4 PHE S 68 ASP S 73 -1 N ASP S 73 O THR S 78 SHEET 1 AB3 6 GLY S 10 VAL S 12 0 SHEET 2 AB3 6 THR S 115 VAL S 119 1 O THR S 116 N GLY S 10 SHEET 3 AB3 6 ALA S 92 SER S 99 -1 N TYR S 94 O THR S 115 SHEET 4 AB3 6 GLY S 33 GLN S 39 -1 N VAL S 37 O TYR S 95 SHEET 5 AB3 6 LEU S 45 ILE S 51 -1 O GLU S 46 N ARG S 38 SHEET 6 AB3 6 ILE S 58 TYR S 60 -1 O TYR S 59 N TYR S 50 SHEET 1 AB4 4 GLY S 10 VAL S 12 0 SHEET 2 AB4 4 THR S 115 VAL S 119 1 O THR S 116 N GLY S 10 SHEET 3 AB4 4 ALA S 92 SER S 99 -1 N TYR S 94 O THR S 115 SHEET 4 AB4 4 PHE S 110 TRP S 111 -1 O PHE S 110 N ARG S 98 SHEET 1 AB5 4 MET S 140 THR S 141 0 SHEET 2 AB5 4 VAL S 155 SER S 161 -1 O ARG S 160 N THR S 141 SHEET 3 AB5 4 ALA S 211 ILE S 216 -1 O PHE S 212 N CYS S 159 SHEET 4 AB5 4 PHE S 203 SER S 208 -1 N SER S 204 O THR S 215 SHEET 1 AB6 6 SER S 146 PRO S 148 0 SHEET 2 AB6 6 THR S 243 GLU S 246 1 O LYS S 244 N VAL S 147 SHEET 3 AB6 6 VAL S 226 GLN S 231 -1 N TYR S 227 O THR S 243 SHEET 4 AB6 6 LEU S 174 GLN S 179 -1 N TYR S 175 O MET S 230 SHEET 5 AB6 6 GLN S 186 TYR S 190 -1 O LEU S 188 N TRP S 176 SHEET 6 AB6 6 ASN S 194 LEU S 195 -1 O ASN S 194 N TYR S 190 SHEET 1 AB7 4 SER S 146 PRO S 148 0 SHEET 2 AB7 4 THR S 243 GLU S 246 1 O LYS S 244 N VAL S 147 SHEET 3 AB7 4 VAL S 226 GLN S 231 -1 N TYR S 227 O THR S 243 SHEET 4 AB7 4 THR S 238 PHE S 239 -1 O THR S 238 N GLN S 231 SSBOND 1 CYS R 9 CYS R 258 1555 1555 2.00 SSBOND 2 CYS R 90 CYS R 168 1555 1555 2.15 SSBOND 3 CYS S 22 CYS S 96 1555 1555 2.02 SSBOND 4 CYS S 159 CYS S 229 1555 1555 2.02 CISPEP 1 PHE R 172 PRO R 173 0 -2.04 CISPEP 2 TYR S 235 PRO S 236 0 5.49 CRYST1 1.000 1.000 1.000 90.00 90.00 90.00 P 1 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