HEADER SIGNALING PROTEIN/IMMUNE SYSTEM 02-DEC-24 9KT9 TITLE CRYO-EM STRUCTURE OF HCA1-GI COMPLEX WITH 3,5-DHBA COMPND MOL_ID: 1; COMPND 2 MOLECULE: GUANINE NUCLEOTIDE-BINDING PROTEIN G(I) SUBUNIT ALPHA-1; COMPND 3 CHAIN: A; COMPND 4 SYNONYM: ADENYLATE CYCLASE-INHIBITING G ALPHA PROTEIN; COMPND 5 ENGINEERED: YES; COMPND 6 MUTATION: YES; COMPND 7 MOL_ID: 2; COMPND 8 MOLECULE: GUANINE NUCLEOTIDE-BINDING PROTEIN G(I)/G(S)/G(T) SUBUNIT COMPND 9 BETA-1; COMPND 10 CHAIN: B; COMPND 11 SYNONYM: TRANSDUCIN BETA CHAIN 1; COMPND 12 ENGINEERED: YES; COMPND 13 MOL_ID: 3; COMPND 14 MOLECULE: GUANINE NUCLEOTIDE-BINDING PROTEIN G(I)/G(S)/G(O) SUBUNIT COMPND 15 GAMMA-2; COMPND 16 CHAIN: C; COMPND 17 SYNONYM: G GAMMA-I; COMPND 18 ENGINEERED: YES; COMPND 19 MOL_ID: 4; COMPND 20 MOLECULE: SCFV16; COMPND 21 CHAIN: E; COMPND 22 ENGINEERED: YES; COMPND 23 MOL_ID: 5; COMPND 24 MOLECULE: HYDROXYCARBOXYLIC ACID RECEPTOR 1; COMPND 25 CHAIN: R; COMPND 26 SYNONYM: G-PROTEIN COUPLED RECEPTOR 104,G-PROTEIN COUPLED RECEPTOR COMPND 27 81; COMPND 28 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: GNAI1; SOURCE 6 EXPRESSION_SYSTEM: SPODOPTERA FRUGIPERDA; SOURCE 7 EXPRESSION_SYSTEM_TAXID: 7108; SOURCE 8 MOL_ID: 2; SOURCE 9 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 10 ORGANISM_COMMON: HUMAN; SOURCE 11 ORGANISM_TAXID: 9606; SOURCE 12 GENE: GNB1; 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: GNG2; SOURCE 20 EXPRESSION_SYSTEM: SPODOPTERA FRUGIPERDA; SOURCE 21 EXPRESSION_SYSTEM_TAXID: 7108; SOURCE 22 MOL_ID: 4; SOURCE 23 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 24 ORGANISM_TAXID: 9606; SOURCE 25 EXPRESSION_SYSTEM: SPODOPTERA FRUGIPERDA; SOURCE 26 EXPRESSION_SYSTEM_TAXID: 7108; SOURCE 27 MOL_ID: 5; SOURCE 28 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 29 ORGANISM_COMMON: HUMAN; SOURCE 30 ORGANISM_TAXID: 9606; SOURCE 31 GENE: HCAR1, GPR104, GPR81, HCA1, FKSG80; SOURCE 32 EXPRESSION_SYSTEM: SPODOPTERA FRUGIPERDA; SOURCE 33 EXPRESSION_SYSTEM_TAXID: 7108 KEYWDS G-PROTEIN-COUPLED RECEPTORS; HYDROXYCARBOXYLIC ACID RECEPTORS, CRYO- KEYWDS 2 EM; SIGNALING PROTEIN, SIGNALING PROTEIN/IMMUNE SYSTEM, SIGNALING KEYWDS 3 PROTEIN-IMMUNE SYSTEM COMPLEX EXPDTA ELECTRON MICROSCOPY AUTHOR S.WU REVDAT 1 10-DEC-25 9KT9 0 JRNL AUTH J.WANG,Y.QIAN,Z.HAN,Y.WANG,Y.LIU,J.LI,Q.DUANMU,S.YE,A.QIAO, JRNL AUTH 2 S.WU JRNL TITL INSIGHTS INTO THE ACTIVATION MECHANISM OF HCA1, HCA2, AND JRNL TITL 2 HCA3. JRNL REF J.MED.CHEM. V. 68 4527 2025 JRNL REFN ISSN 0022-2623 JRNL PMID 39936872 JRNL DOI 10.1021/ACS.JMEDCHEM.4C02567 REMARK 2 REMARK 2 RESOLUTION. 2.70 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 SOFTWARE PACKAGES : NULL 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) : 2.700 REMARK 3 NUMBER OF PARTICLES : 918047 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 9KT9 COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY PDBC ON 02-DEC-24. REMARK 100 THE DEPOSITION ID IS D_1300050747. 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 HCA1-GI REMARK 245 COMPLEX WITH 3,5-DHBA 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 : 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 BIOCONTINUUM (6K X REMARK 245 4K) REMARK 245 MINIMUM DEFOCUS (NM) : 1000.00 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 : NULL REMARK 245 IMAGING MODE : BRIGHT FIELD REMARK 245 ELECTRON DOSE (ELECTRONS NM**-2) : 5400.00 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: A, B, C, E, 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 1 REMARK 465 ILE A 56 REMARK 465 HIS A 57 REMARK 465 GLU A 58 REMARK 465 ALA A 59 REMARK 465 GLY A 60 REMARK 465 TYR A 61 REMARK 465 SER A 62 REMARK 465 GLU A 63 REMARK 465 GLU A 64 REMARK 465 GLU A 65 REMARK 465 CYS A 66 REMARK 465 LYS A 67 REMARK 465 GLN A 68 REMARK 465 TYR A 69 REMARK 465 LYS A 70 REMARK 465 ALA A 71 REMARK 465 VAL A 72 REMARK 465 VAL A 73 REMARK 465 TYR A 74 REMARK 465 SER A 75 REMARK 465 ASN A 76 REMARK 465 THR A 77 REMARK 465 ILE A 78 REMARK 465 GLN A 79 REMARK 465 SER A 80 REMARK 465 ILE A 81 REMARK 465 ILE A 82 REMARK 465 ALA A 83 REMARK 465 ILE A 84 REMARK 465 ILE A 85 REMARK 465 ARG A 86 REMARK 465 ALA A 87 REMARK 465 MET A 88 REMARK 465 GLY A 89 REMARK 465 ARG A 90 REMARK 465 LEU A 91 REMARK 465 LYS A 92 REMARK 465 ILE A 93 REMARK 465 ASP A 94 REMARK 465 PHE A 95 REMARK 465 GLY A 96 REMARK 465 ASP A 97 REMARK 465 SER A 98 REMARK 465 ALA A 99 REMARK 465 ARG A 100 REMARK 465 ALA A 101 REMARK 465 ASP A 102 REMARK 465 ASP A 103 REMARK 465 ALA A 104 REMARK 465 ARG A 105 REMARK 465 GLN A 106 REMARK 465 LEU A 107 REMARK 465 PHE A 108 REMARK 465 VAL A 109 REMARK 465 LEU A 110 REMARK 465 ALA A 111 REMARK 465 GLY A 112 REMARK 465 ALA A 113 REMARK 465 ALA A 114 REMARK 465 GLU A 115 REMARK 465 GLU A 116 REMARK 465 GLY A 117 REMARK 465 PHE A 118 REMARK 465 MET A 119 REMARK 465 THR A 120 REMARK 465 ALA A 121 REMARK 465 GLU A 122 REMARK 465 LEU A 123 REMARK 465 ALA A 124 REMARK 465 GLY A 125 REMARK 465 VAL A 126 REMARK 465 ILE A 127 REMARK 465 LYS A 128 REMARK 465 ARG A 129 REMARK 465 LEU A 130 REMARK 465 TRP A 131 REMARK 465 LYS A 132 REMARK 465 ASP A 133 REMARK 465 SER A 134 REMARK 465 GLY A 135 REMARK 465 VAL A 136 REMARK 465 GLN A 137 REMARK 465 ALA A 138 REMARK 465 CYS A 139 REMARK 465 PHE A 140 REMARK 465 ASN A 141 REMARK 465 ARG A 142 REMARK 465 SER A 143 REMARK 465 ARG A 144 REMARK 465 GLU A 145 REMARK 465 TYR A 146 REMARK 465 GLN A 147 REMARK 465 LEU A 148 REMARK 465 ASN A 149 REMARK 465 ASP A 150 REMARK 465 SER A 151 REMARK 465 ALA A 152 REMARK 465 ALA A 153 REMARK 465 TYR A 154 REMARK 465 TYR A 155 REMARK 465 LEU A 156 REMARK 465 ASN A 157 REMARK 465 ASP A 158 REMARK 465 LEU A 159 REMARK 465 ASP A 160 REMARK 465 ARG A 161 REMARK 465 ILE A 162 REMARK 465 ALA A 163 REMARK 465 GLN A 164 REMARK 465 PRO A 165 REMARK 465 ASN A 166 REMARK 465 TYR A 167 REMARK 465 ILE A 168 REMARK 465 PRO A 169 REMARK 465 THR A 170 REMARK 465 GLN A 171 REMARK 465 GLN A 172 REMARK 465 ASP A 173 REMARK 465 VAL A 174 REMARK 465 LEU A 175 REMARK 465 ARG A 176 REMARK 465 THR A 177 REMARK 465 ARG A 178 REMARK 465 VAL A 179 REMARK 465 LYS A 180 REMARK 465 MET C 1 REMARK 465 ALA C 2 REMARK 465 SER C 3 REMARK 465 ASN C 4 REMARK 465 ASN C 5 REMARK 465 GLU C 63 REMARK 465 LYS C 64 REMARK 465 LYS C 65 REMARK 465 PHE C 66 REMARK 465 PHE C 67 REMARK 465 CYS C 68 REMARK 465 ALA C 69 REMARK 465 ILE C 70 REMARK 465 LEU C 71 REMARK 465 GLY E 122 REMARK 465 GLY E 123 REMARK 465 GLY E 124 REMARK 465 GLY E 125 REMARK 465 SER E 126 REMARK 465 GLY E 127 REMARK 465 GLY E 128 REMARK 465 GLY E 129 REMARK 465 GLY E 130 REMARK 465 SER E 131 REMARK 465 GLY E 132 REMARK 465 GLY E 133 REMARK 465 GLY E 134 REMARK 465 GLY E 135 REMARK 465 LYS E 248 REMARK 465 ALA E 249 REMARK 465 ALA E 250 REMARK 465 ALA E 251 REMARK 465 LEU E 252 REMARK 465 GLU E 253 REMARK 465 VAL E 254 REMARK 465 LEU E 255 REMARK 465 PHE E 256 REMARK 465 GLN E 257 REMARK 465 GLY E 258 REMARK 465 PRO E 259 REMARK 465 HIS E 260 REMARK 465 HIS E 261 REMARK 465 HIS E 262 REMARK 465 HIS E 263 REMARK 465 HIS E 264 REMARK 465 HIS E 265 REMARK 465 HIS E 266 REMARK 465 HIS E 267 REMARK 465 MET R 1 REMARK 465 TYR R 2 REMARK 465 ASN R 3 REMARK 465 GLY R 4 REMARK 465 SER R 5 REMARK 465 ASN R 290 REMARK 465 LYS R 291 REMARK 465 LEU R 292 REMARK 465 LYS R 293 REMARK 465 ILE R 294 REMARK 465 CYS R 295 REMARK 465 SER R 296 REMARK 465 LEU R 297 REMARK 465 LYS R 298 REMARK 465 PRO R 299 REMARK 465 LYS R 300 REMARK 465 GLN R 301 REMARK 465 PRO R 302 REMARK 465 GLY R 303 REMARK 465 HIS R 304 REMARK 465 SER R 305 REMARK 465 LYS R 306 REMARK 465 THR R 307 REMARK 465 GLN R 308 REMARK 465 ARG R 309 REMARK 465 PRO R 310 REMARK 465 GLU R 311 REMARK 465 GLU R 312 REMARK 465 MET R 313 REMARK 465 PRO R 314 REMARK 465 ILE R 315 REMARK 465 SER R 316 REMARK 465 ASN R 317 REMARK 465 LEU R 318 REMARK 465 GLY R 319 REMARK 465 ARG R 320 REMARK 465 ARG R 321 REMARK 465 SER R 322 REMARK 465 CYS R 323 REMARK 465 ILE R 324 REMARK 465 SER R 325 REMARK 465 VAL R 326 REMARK 465 ALA R 327 REMARK 465 ASN R 328 REMARK 465 SER R 329 REMARK 465 PHE R 330 REMARK 465 GLN R 331 REMARK 465 SER R 332 REMARK 465 GLN R 333 REMARK 465 SER R 334 REMARK 465 ASP R 335 REMARK 465 GLY R 336 REMARK 465 GLN R 337 REMARK 465 TRP R 338 REMARK 465 ASP R 339 REMARK 465 PRO R 340 REMARK 465 HIS R 341 REMARK 465 ILE R 342 REMARK 465 VAL R 343 REMARK 465 GLU R 344 REMARK 465 TRP R 345 REMARK 465 HIS R 346 REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: TORSION ANGLES REMARK 500 REMARK 500 TORSION ANGLES OUTSIDE THE EXPECTED RAMACHANDRAN REGIONS: REMARK 500 (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN IDENTIFIER; REMARK 500 SSEQ=SEQUENCE NUMBER; I=INSERTION CODE). REMARK 500 REMARK 500 STANDARD TABLE: REMARK 500 FORMAT:(10X,I3,1X,A3,1X,A1,I4,A1,4X,F7.2,3X,F7.2) REMARK 500 REMARK 500 EXPECTED VALUES: GJ KLEYWEGT AND TA JONES (1996). PHI/PSI- REMARK 500 CHOLOGY: RAMACHANDRAN REVISITED. STRUCTURE 4, 1395 - 1400 REMARK 500 REMARK 500 M RES CSSEQI PSI PHI REMARK 500 LYS A 314 -66.81 61.61 REMARK 500 SER A 326 31.37 -143.13 REMARK 500 ALA B 28 -171.27 58.06 REMARK 500 TRP B 99 52.45 -90.70 REMARK 500 MET B 101 29.33 -143.28 REMARK 500 SER B 334 31.64 72.46 REMARK 500 VAL E 48 -61.10 -120.18 REMARK 500 ALA E 92 -175.88 -170.65 REMARK 500 MET E 192 -12.79 73.84 REMARK 500 ASP E 223 51.65 -90.25 REMARK 500 PRO R 49 172.32 -59.92 REMARK 500 SER R 50 -5.25 72.53 REMARK 500 PRO R 87 -6.39 -59.28 REMARK 500 THR R 94 46.24 -88.04 REMARK 500 ILE R 169 -60.95 -93.04 REMARK 500 GLU R 171 -129.27 70.03 REMARK 500 HIS R 177 -4.07 65.56 REMARK 500 ASP R 178 -56.22 -120.30 REMARK 500 THR R 246 47.74 -93.74 REMARK 500 SER R 281 53.17 -92.02 REMARK 500 REMARK 500 REMARK: NULL REMARK 900 REMARK 900 RELATED ENTRIES REMARK 900 RELATED ID: EMD-62560 RELATED DB: EMDB REMARK 900 CRYO-EM STRUCTURE OF HCA1-GI COMPLEX WITH 3,5-DHBA DBREF 9KT9 A 1 354 UNP P63096 GNAI1_HUMAN 1 354 DBREF 9KT9 B 2 340 UNP P62873 GBB1_HUMAN 2 340 DBREF 9KT9 C 1 71 UNP P59768 GBG2_HUMAN 1 71 DBREF 9KT9 E 1 267 PDB 9KT9 9KT9 1 267 DBREF 9KT9 R 1 346 UNP Q9BXC0 HCAR1_HUMAN 1 346 SEQADV 9KT9 ASN A 47 UNP P63096 SER 47 ENGINEERED MUTATION SEQADV 9KT9 ALA A 203 UNP P63096 GLY 203 ENGINEERED MUTATION SEQADV 9KT9 ALA A 245 UNP P63096 GLU 245 ENGINEERED MUTATION SEQADV 9KT9 SER A 326 UNP P63096 ALA 326 ENGINEERED MUTATION SEQADV 9KT9 GLN B 1 UNP P62873 EXPRESSION TAG SEQRES 1 A 354 MET GLY CYS THR LEU SER ALA GLU ASP LYS ALA ALA VAL SEQRES 2 A 354 GLU ARG SER LYS MET ILE ASP ARG ASN LEU ARG GLU ASP SEQRES 3 A 354 GLY GLU LYS ALA ALA ARG GLU VAL LYS LEU LEU LEU LEU SEQRES 4 A 354 GLY ALA GLY GLU SER GLY LYS ASN THR ILE VAL LYS GLN SEQRES 5 A 354 MET LYS ILE ILE HIS GLU ALA GLY TYR SER GLU GLU GLU SEQRES 6 A 354 CYS LYS GLN TYR LYS ALA VAL VAL TYR SER ASN THR ILE SEQRES 7 A 354 GLN SER ILE ILE ALA ILE ILE ARG ALA MET GLY ARG LEU SEQRES 8 A 354 LYS ILE ASP PHE GLY ASP SER ALA ARG ALA ASP ASP ALA SEQRES 9 A 354 ARG GLN LEU PHE VAL LEU ALA GLY ALA ALA GLU GLU GLY SEQRES 10 A 354 PHE MET THR ALA GLU LEU ALA GLY VAL ILE LYS ARG LEU SEQRES 11 A 354 TRP LYS ASP SER GLY VAL GLN ALA CYS PHE ASN ARG SER SEQRES 12 A 354 ARG GLU TYR GLN LEU ASN ASP SER ALA ALA TYR TYR LEU SEQRES 13 A 354 ASN ASP LEU ASP ARG ILE ALA GLN PRO ASN TYR ILE PRO SEQRES 14 A 354 THR GLN GLN ASP VAL LEU ARG THR ARG VAL LYS THR THR SEQRES 15 A 354 GLY ILE VAL GLU THR HIS PHE THR PHE LYS ASP LEU HIS SEQRES 16 A 354 PHE LYS MET PHE ASP VAL GLY ALA GLN ARG SER GLU ARG SEQRES 17 A 354 LYS LYS TRP ILE HIS CYS PHE GLU GLY VAL THR ALA ILE SEQRES 18 A 354 ILE PHE CYS VAL ALA LEU SER ASP TYR ASP LEU VAL LEU SEQRES 19 A 354 ALA GLU ASP GLU GLU MET ASN ARG MET HIS ALA SER MET SEQRES 20 A 354 LYS LEU PHE ASP SER ILE CYS ASN ASN LYS TRP PHE THR SEQRES 21 A 354 ASP THR SER ILE ILE LEU PHE LEU ASN LYS LYS ASP LEU SEQRES 22 A 354 PHE GLU GLU LYS ILE LYS LYS SER PRO LEU THR ILE CYS SEQRES 23 A 354 TYR PRO GLU TYR ALA GLY SER ASN THR TYR GLU GLU ALA SEQRES 24 A 354 ALA ALA TYR ILE GLN CYS GLN PHE GLU ASP LEU ASN LYS SEQRES 25 A 354 ARG LYS ASP THR LYS GLU ILE TYR THR HIS PHE THR CYS SEQRES 26 A 354 SER THR ASP THR LYS ASN VAL GLN PHE VAL PHE ASP ALA SEQRES 27 A 354 VAL THR ASP VAL ILE ILE LYS ASN ASN LEU LYS ASP CYS SEQRES 28 A 354 GLY LEU PHE SEQRES 1 B 340 GLN SER GLU LEU ASP GLN LEU ARG GLN GLU ALA GLU GLN SEQRES 2 B 340 LEU LYS ASN GLN ILE ARG ASP ALA ARG LYS ALA CYS ALA SEQRES 3 B 340 ASP ALA THR LEU SER GLN ILE THR ASN ASN ILE ASP PRO SEQRES 4 B 340 VAL GLY ARG ILE GLN MET ARG THR ARG ARG THR LEU ARG SEQRES 5 B 340 GLY HIS LEU ALA LYS ILE TYR ALA MET HIS TRP GLY THR SEQRES 6 B 340 ASP SER ARG LEU LEU VAL SER ALA SER GLN ASP GLY LYS SEQRES 7 B 340 LEU ILE ILE TRP ASP SER TYR THR THR ASN LYS VAL HIS SEQRES 8 B 340 ALA ILE PRO LEU ARG SER SER TRP VAL MET THR CYS ALA SEQRES 9 B 340 TYR ALA PRO SER GLY ASN TYR VAL ALA CYS GLY GLY LEU SEQRES 10 B 340 ASP ASN ILE CYS SER ILE TYR ASN LEU LYS THR ARG GLU SEQRES 11 B 340 GLY ASN VAL ARG VAL SER ARG GLU LEU ALA GLY HIS THR SEQRES 12 B 340 GLY TYR LEU SER CYS CYS ARG PHE LEU ASP ASP ASN GLN SEQRES 13 B 340 ILE VAL THR SER SER GLY ASP THR THR CYS ALA LEU TRP SEQRES 14 B 340 ASP ILE GLU THR GLY GLN GLN THR THR THR PHE THR GLY SEQRES 15 B 340 HIS THR GLY ASP VAL MET SER LEU SER LEU ALA PRO ASP SEQRES 16 B 340 THR ARG LEU PHE VAL SER GLY ALA CYS ASP ALA SER ALA SEQRES 17 B 340 LYS LEU TRP ASP VAL ARG GLU GLY MET CYS ARG GLN THR SEQRES 18 B 340 PHE THR GLY HIS GLU SER ASP ILE ASN ALA ILE CYS PHE SEQRES 19 B 340 PHE PRO ASN GLY ASN ALA PHE ALA THR GLY SER ASP ASP SEQRES 20 B 340 ALA THR CYS ARG LEU PHE ASP LEU ARG ALA ASP GLN GLU SEQRES 21 B 340 LEU MET THR TYR SER HIS ASP ASN ILE ILE CYS GLY ILE SEQRES 22 B 340 THR SER VAL SER PHE SER LYS SER GLY ARG LEU LEU LEU SEQRES 23 B 340 ALA GLY TYR ASP ASP PHE ASN CYS ASN VAL TRP ASP ALA SEQRES 24 B 340 LEU LYS ALA ASP ARG ALA GLY VAL LEU ALA GLY HIS ASP SEQRES 25 B 340 ASN ARG VAL SER CYS LEU GLY VAL THR ASP ASP GLY MET SEQRES 26 B 340 ALA VAL ALA THR GLY SER TRP ASP SER PHE LEU LYS ILE SEQRES 27 B 340 TRP ASN SEQRES 1 C 71 MET ALA SER ASN ASN THR ALA SER ILE ALA GLN ALA ARG SEQRES 2 C 71 LYS LEU VAL GLU GLN LEU LYS MET GLU ALA ASN ILE ASP SEQRES 3 C 71 ARG ILE LYS VAL SER LYS ALA ALA ALA ASP LEU MET ALA SEQRES 4 C 71 TYR CYS GLU ALA HIS ALA LYS GLU ASP PRO LEU LEU THR SEQRES 5 C 71 PRO VAL PRO ALA SER GLU ASN PRO PHE ARG GLU LYS LYS SEQRES 6 C 71 PHE PHE CYS ALA ILE LEU SEQRES 1 E 267 MET VAL GLN LEU VAL GLU SER GLY GLY GLY LEU VAL GLN SEQRES 2 E 267 PRO GLY GLY SER ARG LYS LEU SER CYS SER ALA SER GLY SEQRES 3 E 267 PHE ALA PHE SER SER PHE GLY MET HIS TRP VAL ARG GLN SEQRES 4 E 267 ALA PRO GLU LYS GLY LEU GLU TRP VAL ALA TYR ILE SER SEQRES 5 E 267 SER GLY SER GLY THR ILE TYR TYR ALA ASP THR VAL LYS SEQRES 6 E 267 GLY ARG PHE THR ILE SER ARG ASP ASP PRO LYS ASN THR SEQRES 7 E 267 LEU PHE LEU GLN MET THR SER LEU ARG SER GLU ASP THR SEQRES 8 E 267 ALA MET TYR TYR CYS VAL ARG SER ILE TYR TYR TYR GLY SEQRES 9 E 267 SER SER PRO PHE ASP PHE TRP GLY GLN GLY THR THR LEU SEQRES 10 E 267 THR VAL SER SER GLY GLY GLY GLY SER GLY GLY GLY GLY SEQRES 11 E 267 SER GLY GLY GLY GLY SER ASP ILE VAL MET THR GLN ALA SEQRES 12 E 267 THR SER SER VAL PRO VAL THR PRO GLY GLU SER VAL SER SEQRES 13 E 267 ILE SER CYS ARG SER SER LYS SER LEU LEU HIS SER ASN SEQRES 14 E 267 GLY ASN THR TYR LEU TYR TRP PHE LEU GLN ARG PRO GLY SEQRES 15 E 267 GLN SER PRO GLN LEU LEU ILE TYR ARG MET SER ASN LEU SEQRES 16 E 267 ALA SER GLY VAL PRO ASP ARG PHE SER GLY SER GLY SER SEQRES 17 E 267 GLY THR ALA PHE THR LEU THR ILE SER ARG LEU GLU ALA SEQRES 18 E 267 GLU ASP VAL GLY VAL TYR TYR CYS MET GLN HIS LEU GLU SEQRES 19 E 267 TYR PRO LEU THR PHE GLY ALA GLY THR LYS LEU GLU LEU SEQRES 20 E 267 LYS ALA ALA ALA LEU GLU VAL LEU PHE GLN GLY PRO HIS SEQRES 21 E 267 HIS HIS HIS HIS HIS HIS HIS SEQRES 1 R 346 MET TYR ASN GLY SER CYS CYS ARG ILE GLU GLY ASP THR SEQRES 2 R 346 ILE SER GLN VAL MET PRO PRO LEU LEU ILE VAL ALA PHE SEQRES 3 R 346 VAL LEU GLY ALA LEU GLY ASN GLY VAL ALA LEU CYS GLY SEQRES 4 R 346 PHE CYS PHE HIS MET LYS THR TRP LYS PRO SER THR VAL SEQRES 5 R 346 TYR LEU PHE ASN LEU ALA VAL ALA ASP PHE LEU LEU MET SEQRES 6 R 346 ILE CYS LEU PRO PHE ARG THR ASP TYR TYR LEU ARG ARG SEQRES 7 R 346 ARG HIS TRP ALA PHE GLY ASP ILE PRO CYS ARG VAL GLY SEQRES 8 R 346 LEU PHE THR LEU ALA MET ASN ARG ALA GLY SER ILE VAL SEQRES 9 R 346 PHE LEU THR VAL VAL ALA ALA ASP ARG TYR PHE LYS VAL SEQRES 10 R 346 VAL HIS PRO HIS HIS ALA VAL ASN THR ILE SER THR ARG SEQRES 11 R 346 VAL ALA ALA GLY ILE VAL CYS THR LEU TRP ALA LEU VAL SEQRES 12 R 346 ILE LEU GLY THR VAL TYR LEU LEU LEU GLU ASN HIS LEU SEQRES 13 R 346 CYS VAL GLN GLU THR ALA VAL SER CYS GLU SER PHE ILE SEQRES 14 R 346 MET GLU SER ALA ASN GLY TRP HIS ASP ILE MET PHE GLN SEQRES 15 R 346 LEU GLU PHE PHE MET PRO LEU GLY ILE ILE LEU PHE CYS SEQRES 16 R 346 SER PHE LYS ILE VAL TRP SER LEU ARG ARG ARG GLN GLN SEQRES 17 R 346 LEU ALA ARG GLN ALA ARG MET LYS LYS ALA THR ARG PHE SEQRES 18 R 346 ILE MET VAL VAL ALA ILE VAL PHE ILE THR CYS TYR LEU SEQRES 19 R 346 PRO SER VAL SER ALA ARG LEU TYR PHE LEU TRP THR VAL SEQRES 20 R 346 PRO SER SER ALA CYS ASP PRO SER VAL HIS GLY ALA LEU SEQRES 21 R 346 HIS ILE THR LEU SER PHE THR TYR MET ASN SER MET LEU SEQRES 22 R 346 ASP PRO LEU VAL TYR TYR PHE SER SER PRO SER PHE PRO SEQRES 23 R 346 LYS PHE TYR ASN LYS LEU LYS ILE CYS SER LEU LYS PRO SEQRES 24 R 346 LYS GLN PRO GLY HIS SER LYS THR GLN ARG PRO GLU GLU SEQRES 25 R 346 MET PRO ILE SER ASN LEU GLY ARG ARG SER CYS ILE SER SEQRES 26 R 346 VAL ALA ASN SER PHE GLN SER GLN SER ASP GLY GLN TRP SEQRES 27 R 346 ASP PRO HIS ILE VAL GLU TRP HIS HET 34D R 401 11 HETNAM 34D 3,5-DIHYDROXYBENZOATE FORMUL 6 34D C7 H6 O4 HELIX 1 AA1 SER A 6 ALA A 31 1 26 HELIX 2 AA2 GLY A 45 LYS A 54 1 10 HELIX 3 AA3 GLU A 207 GLU A 216 5 10 HELIX 4 AA4 ALA A 226 ASP A 231 5 6 HELIX 5 AA5 ASN A 241 ASN A 255 1 15 HELIX 6 AA6 LYS A 271 SER A 281 1 11 HELIX 7 AA7 PRO A 282 CYS A 286 5 5 HELIX 8 AA8 THR A 295 ASP A 309 1 15 HELIX 9 AA9 LYS A 330 GLY A 352 1 23 HELIX 10 AB1 SER B 2 CYS B 25 1 24 HELIX 11 AB2 THR B 29 ASN B 35 1 7 HELIX 12 AB3 SER C 8 ALA C 23 1 16 HELIX 13 AB4 LYS C 29 HIS C 44 1 16 HELIX 14 AB5 ALA C 45 ASP C 48 5 4 HELIX 15 AB6 ALA E 28 PHE E 32 5 5 HELIX 16 AB7 ARG E 87 THR E 91 5 5 HELIX 17 AB8 GLY R 11 HIS R 43 1 33 HELIX 18 AB9 THR R 51 LEU R 68 1 18 HELIX 19 AC1 PRO R 69 THR R 72 5 4 HELIX 20 AC2 ILE R 103 ARG R 113 1 11 HELIX 21 AC3 HIS R 122 ILE R 127 5 6 HELIX 22 AC4 SER R 128 THR R 147 1 20 HELIX 23 AC5 ASP R 178 ARG R 205 1 28 HELIX 24 AC6 GLN R 212 THR R 246 1 35 HELIX 25 AC7 ASP R 253 TYR R 268 1 16 HELIX 26 AC8 MET R 269 TYR R 279 1 11 SHEET 1 AA1 6 VAL A 185 PHE A 191 0 SHEET 2 AA1 6 LEU A 194 ASP A 200 -1 O PHE A 196 N PHE A 189 SHEET 3 AA1 6 VAL A 34 LEU A 39 1 N VAL A 34 O LYS A 197 SHEET 4 AA1 6 ALA A 220 VAL A 225 1 O ILE A 222 N LEU A 39 SHEET 5 AA1 6 SER A 263 LEU A 268 1 O ILE A 265 N ILE A 221 SHEET 6 AA1 6 ILE A 319 PHE A 323 1 O TYR A 320 N LEU A 266 SHEET 1 AA2 4 ARG B 46 LEU B 51 0 SHEET 2 AA2 4 LEU B 336 ASN B 340 -1 O ILE B 338 N ARG B 48 SHEET 3 AA2 4 VAL B 327 SER B 331 -1 N VAL B 327 O TRP B 339 SHEET 4 AA2 4 VAL B 315 VAL B 320 -1 N GLY B 319 O ALA B 328 SHEET 1 AA3 4 ILE B 58 TRP B 63 0 SHEET 2 AA3 4 LEU B 69 SER B 74 -1 O ALA B 73 N ALA B 60 SHEET 3 AA3 4 LYS B 78 ASP B 83 -1 O TRP B 82 N LEU B 70 SHEET 4 AA3 4 LYS B 89 PRO B 94 -1 O ILE B 93 N LEU B 79 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 CYS B 121 ASN B 125 -1 O TYR B 124 N VAL B 112 SHEET 4 AA4 4 ARG B 134 LEU B 139 -1 O ARG B 134 N ASN B 125 SHEET 1 AA5 4 LEU B 146 ASP B 153 0 SHEET 2 AA5 4 GLN B 156 SER B 161 -1 O GLN B 156 N LEU B 152 SHEET 3 AA5 4 THR B 165 ASP B 170 -1 O TRP B 169 N ILE B 157 SHEET 4 AA5 4 GLN B 175 THR B 181 -1 O PHE B 180 N CYS B 166 SHEET 1 AA6 4 VAL B 187 LEU B 192 0 SHEET 2 AA6 4 LEU B 198 ALA B 203 -1 O GLY B 202 N MET B 188 SHEET 3 AA6 4 ALA B 208 ASP B 212 -1 O TRP B 211 N PHE B 199 SHEET 4 AA6 4 MET B 217 PHE B 222 -1 O PHE B 222 N ALA B 208 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 ASN B 230 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 PHE B 278 0 SHEET 2 AA8 4 LEU B 284 TYR B 289 -1 O GLY B 288 N SER B 275 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 LEU B 308 N CYS B 294 SHEET 1 AA9 4 GLN E 3 SER E 7 0 SHEET 2 AA9 4 ARG E 18 SER E 25 -1 O SER E 23 N VAL E 5 SHEET 3 AA9 4 THR E 78 MET E 83 -1 O MET E 83 N ARG E 18 SHEET 4 AA9 4 PHE E 68 ASP E 73 -1 N SER E 71 O PHE E 80 SHEET 1 AB1 6 GLY E 10 VAL E 12 0 SHEET 2 AB1 6 THR E 115 VAL E 119 1 O THR E 116 N GLY E 10 SHEET 3 AB1 6 ALA E 92 SER E 99 -1 N ALA E 92 O LEU E 117 SHEET 4 AB1 6 GLY E 33 GLN E 39 -1 N VAL E 37 O TYR E 95 SHEET 5 AB1 6 LEU E 45 ILE E 51 -1 O GLU E 46 N ARG E 38 SHEET 6 AB1 6 ILE E 58 TYR E 60 -1 O TYR E 59 N TYR E 50 SHEET 1 AB2 4 GLY E 10 VAL E 12 0 SHEET 2 AB2 4 THR E 115 VAL E 119 1 O THR E 116 N GLY E 10 SHEET 3 AB2 4 ALA E 92 SER E 99 -1 N ALA E 92 O LEU E 117 SHEET 4 AB2 4 PHE E 110 TRP E 111 -1 O PHE E 110 N ARG E 98 SHEET 1 AB3 4 MET E 140 THR E 141 0 SHEET 2 AB3 4 VAL E 155 SER E 161 -1 O ARG E 160 N THR E 141 SHEET 3 AB3 4 ALA E 211 ILE E 216 -1 O LEU E 214 N ILE E 157 SHEET 4 AB3 4 PHE E 203 SER E 208 -1 N SER E 206 O THR E 213 SHEET 1 AB4 6 SER E 146 PRO E 148 0 SHEET 2 AB4 6 THR E 243 GLU E 246 1 O LYS E 244 N VAL E 147 SHEET 3 AB4 6 VAL E 226 GLN E 231 -1 N TYR E 227 O THR E 243 SHEET 4 AB4 6 LEU E 174 GLN E 179 -1 N PHE E 177 O TYR E 228 SHEET 5 AB4 6 GLN E 186 TYR E 190 -1 O ILE E 189 N TRP E 176 SHEET 6 AB4 6 ASN E 194 LEU E 195 -1 O ASN E 194 N TYR E 190 SHEET 1 AB5 2 LEU R 156 VAL R 158 0 SHEET 2 AB5 2 VAL R 163 CYS R 165 -1 O SER R 164 N CYS R 157 SSBOND 1 CYS E 159 CYS E 229 1555 1555 2.04 SSBOND 2 CYS R 6 CYS R 157 1555 1555 2.03 SSBOND 3 CYS R 7 CYS R 252 1555 1555 2.03 SSBOND 4 CYS R 88 CYS R 165 1555 1555 2.03 CISPEP 1 TYR E 235 PRO E 236 0 0.07 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