HEADER MEMBRANE PROTEIN 03-APR-23 8SBB TITLE CRYO-EM STRUCTURE OF FTALKB COMPND MOL_ID: 1; COMPND 2 MOLECULE: ALKANE 1-MONOOXYGENASE; COMPND 3 CHAIN: A; COMPND 4 ENGINEERED: YES; COMPND 5 MOL_ID: 2; COMPND 6 MOLECULE: NANOBODY; COMPND 7 CHAIN: B; COMPND 8 ENGINEERED: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: FONTIMONAS THERMOPHILA; SOURCE 3 ORGANISM_TAXID: 1076937; SOURCE 4 GENE: SAMN04488120_1209; SOURCE 5 EXPRESSION_SYSTEM: ESCHERICHIA COLI BL21(DE3); SOURCE 6 EXPRESSION_SYSTEM_TAXID: 469008; SOURCE 7 MOL_ID: 2; SOURCE 8 ORGANISM_SCIENTIFIC: SYNTHETIC CONSTRUCT; SOURCE 9 ORGANISM_TAXID: 32630; SOURCE 10 EXPRESSION_SYSTEM: ESCHERICHIA COLI BL21(DE3); SOURCE 11 EXPRESSION_SYSTEM_TAXID: 469008 KEYWDS ENZYME, MEMBRANE PROTEIN EXPDTA ELECTRON MICROSCOPY AUTHOR J.ZHANG,L.FENG REVDAT 1 26-APR-23 8SBB 0 JRNL AUTH X.GUO,J.ZHANG,L.HAN,J.LEE,S.C.WILLIAMS,A.FORSBERG,Y.XU, JRNL AUTH 2 R.N.AUSTIN,L.FENG JRNL TITL STRUCTURE AND MECHANISM OF THE ALKANE-OXIDIZING ENZYME ALKB. JRNL REF NAT COMMUN V. 14 2180 2023 JRNL REFN ESSN 2041-1723 JRNL PMID 37069165 JRNL DOI 10.1038/S41467-023-37869-Z REMARK 2 REMARK 2 RESOLUTION. 3.59 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) : 3.590 REMARK 3 NUMBER OF PARTICLES : 107776 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 8SBB COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 03-APR-23. REMARK 100 THE DEPOSITION ID IS D_1000272175. 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 : FTAKLB-NANOBODY 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 : 7.00 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 : FEI TITAN KRIOS REMARK 245 DETECTOR TYPE : FEI FALCON IV (4K X 4K) REMARK 245 MINIMUM DEFOCUS (NM) : 1600.00 REMARK 245 MAXIMUM DEFOCUS (NM) : 2200.00 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: DIMERIC REMARK 350 APPLY THE FOLLOWING TO CHAINS: A, B 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 ALA A 2 REMARK 465 THR A 3 REMARK 465 SER A 4 REMARK 465 MET A 5 REMARK 465 GLY A 6 REMARK 465 SER A 7 REMARK 465 HIS A 8 REMARK 465 PRO A 9 REMARK 465 GLY A 10 REMARK 465 PHE A 11 REMARK 465 GLY A 12 REMARK 465 ALA A 13 REMARK 465 ASP A 14 REMARK 465 SER A 15 REMARK 465 SER A 16 REMARK 465 GLY A 17 REMARK 465 THR A 18 REMARK 465 GLY A 399 REMARK 465 MET B 1 REMARK 465 ALA B 2 REMARK 465 VAL B 122 REMARK 465 SER B 123 REMARK 465 SER B 124 REMARK 465 HIS B 125 REMARK 465 HIS B 126 REMARK 465 HIS B 127 REMARK 465 HIS B 128 REMARK 465 HIS B 129 REMARK 465 HIS B 130 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 B 3 CG CD OE1 NE2 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 TYR A 21 54.11 -106.46 REMARK 500 ASP A 23 -179.52 -62.42 REMARK 500 ARG A 24 -35.31 -132.97 REMARK 500 PHE A 54 0.32 -65.57 REMARK 500 PRO A 81 -175.09 -67.13 REMARK 500 GLU A 89 40.71 -101.06 REMARK 500 HIS A 119 43.22 -140.35 REMARK 500 HIS A 151 47.15 -68.54 REMARK 500 ARG A 225 -8.92 -55.39 REMARK 500 SER A 229 154.72 -47.20 REMARK 500 VAL A 257 0.63 -66.20 REMARK 500 ASP A 292 14.02 58.02 REMARK 500 TYR A 301 39.07 -94.74 REMARK 500 HIS A 317 107.89 -42.05 REMARK 500 GLN A 319 8.04 -68.89 REMARK 500 TYR A 332 43.36 -81.22 REMARK 500 ARG A 336 -168.12 -78.60 REMARK 500 VAL A 364 -64.77 -108.80 REMARK 500 GLN A 386 32.96 -143.84 REMARK 500 ALA B 26 -165.86 -79.67 REMARK 500 ARG B 32 17.94 -150.00 REMARK 500 TRP B 34 -165.30 -176.45 REMARK 500 ARG B 73 -122.08 -116.70 REMARK 500 ASP B 74 45.03 77.81 REMARK 500 ALA B 76 154.18 -48.06 REMARK 500 PRO B 104 137.95 -39.68 REMARK 500 REMARK 500 REMARK: NULL REMARK 620 REMARK 620 METAL COORDINATION REMARK 620 (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN IDENTIFIER; REMARK 620 SSEQ=SEQUENCE NUMBER; I=INSERTION CODE): REMARK 620 REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL REMARK 620 FE A 402 FE REMARK 620 N RES CSSEQI ATOM REMARK 620 1 HIS A 147 NE2 REMARK 620 2 HIS A 151 NE2 80.1 REMARK 620 3 HIS A 177 NE2 81.2 88.2 REMARK 620 4 HIS A 182 NE2 163.8 93.1 83.9 REMARK 620 5 HIS A 324 NE2 96.6 102.8 168.3 99.3 REMARK 620 N 1 2 3 4 REMARK 620 REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL REMARK 620 FE A 401 FE REMARK 620 N RES CSSEQI ATOM REMARK 620 1 HIS A 181 NE2 REMARK 620 2 HIS A 282 NE2 95.0 REMARK 620 3 HIS A 321 NE2 160.3 99.1 REMARK 620 4 HIS A 325 NE2 89.8 106.2 99.3 REMARK 620 N 1 2 3 REMARK 900 REMARK 900 RELATED ENTRIES REMARK 900 RELATED ID: EMD-40303 RELATED DB: EMDB REMARK 900 CRYO-EM STRUCTURE OF FTALKB DBREF1 8SBB A 1 399 UNP A0A1I2KHB9_9GAMM DBREF2 8SBB A A0A1I2KHB9 1 399 DBREF 8SBB B 1 130 PDB 8SBB 8SBB 1 130 SEQRES 1 A 399 MET ALA THR SER MET GLY SER HIS PRO GLY PHE GLY ALA SEQRES 2 A 399 ASP SER SER GLY THR ILE ALA TYR ARG ASP ARG LYS ARG SEQRES 3 A 399 PRO PHE TRP THR LEU SER VAL LEU TRP PRO VAL SER PRO SEQRES 4 A 399 LEU ILE GLY ILE TYR LEU ALA HIS THR THR GLY VAL GLY SEQRES 5 A 399 ALA PHE PHE TRP LEU THR LEU ALA VAL TRP TYR LEU ILE SEQRES 6 A 399 ILE PRO LEU LEU ASP TRP VAL LEU GLY ASP ASP GLN SER SEQRES 7 A 399 ASN PRO PRO GLU ALA VAL VAL PRO ALA LEU GLU SER ASP SEQRES 8 A 399 ARG TYR TYR ARG ILE LEU THR TYR LEU THR VAL PRO ILE SEQRES 9 A 399 HIS TYR VAL VAL LEU ILE GLY SER ALA TRP TYR VAL SER SEQRES 10 A 399 THR HIS TYR GLY SER MET THR TRP TYR ASP ILE LEU GLY SEQRES 11 A 399 LEU ALA LEU SER VAL GLY ILE VAL ASN GLY LEU ALA ILE SEQRES 12 A 399 ASN THR GLY HIS GLU LEU GLY HIS LYS LYS THR GLU LEU SEQRES 13 A 399 GLU ARG TRP LEU ALA LYS ILE VAL LEU ALA VAL VAL GLY SEQRES 14 A 399 TYR GLY HIS PHE PHE ILE GLU HIS ASN LYS GLY HIS HIS SEQRES 15 A 399 LYS ASP VAL ALA THR PRO GLU ASP PRO ALA SER ALA PRO SEQRES 16 A 399 PHE GLY GLN SER ILE TYR ARG PHE ALA LEU ARG GLU ILE SEQRES 17 A 399 PRO GLY ALA ILE ILE ARG ALA TRP ASN SER GLU LYS GLU SEQRES 18 A 399 ARG LEU GLY ARG LEU GLY HIS SER PRO TRP SER LEU GLN SEQRES 19 A 399 ASN GLU VAL LEU GLN PRO LEU LEU ILE THR ILE PRO LEU SEQRES 20 A 399 TYR VAL GLY LEU ILE ALA VAL LEU GLY VAL LYS ILE ILE SEQRES 21 A 399 PRO PHE LEU LEU ILE GLN ILE VAL PHE GLY TRP TRP GLN SEQRES 22 A 399 LEU THR SER ALA ASN TYR ILE GLU HIS TYR GLY LEU LEU SEQRES 23 A 399 ARG GLN LYS LEU PRO ASP GLY ARG TYR GLU ARG CYS GLN SEQRES 24 A 399 PRO TYR HIS SER TRP ASN SER ASN HIS VAL MET SER ASN SEQRES 25 A 399 LEU ILE LEU PHE HIS LEU GLN ARG HIS SER ASP HIS HIS SEQRES 26 A 399 ALA HIS PRO THR ARG ARG TYR GLN SER LEU ARG ASP PHE SEQRES 27 A 399 PRO ASP LEU PRO THR LEU PRO SER GLY TYR PRO LEU MET SEQRES 28 A 399 PHE ALA LEU SER TYR PHE PRO PRO LEU TRP ARG ALA VAL SEQRES 29 A 399 MET ASP ARG ARG VAL LEU ASP VAL CYS GLY ARG ASP ALA SEQRES 30 A 399 ARG LYS ILE ASN ILE ASP PRO ASN GLN ARG GLU LYS ILE SEQRES 31 A 399 ILE HIS LYS PHE ASN LEU LEU THR GLY SEQRES 1 B 130 MET ALA GLN VAL GLN LEU GLN GLU SER GLY GLY GLY LEU SEQRES 2 B 130 VAL GLN ALA GLY GLY SER LEU ARG LEU SER CYS ALA ALA SEQRES 3 B 130 SER GLY THR ILE SER ARG TYR TRP THR MET GLY TRP TYR SEQRES 4 B 130 ARG GLN ALA PRO GLY LYS GLU ARG GLU LEU VAL ALA GLY SEQRES 5 B 130 ILE SER GLU GLY GLY SER THR ASN TYR ALA ASP SER VAL SEQRES 6 B 130 LYS GLY ARG PHE THR ILE SER ARG ASP ASN ALA LYS ASN SEQRES 7 B 130 THR VAL TYR LEU GLN MET ASN SER LEU LYS PRO GLU ASP SEQRES 8 B 130 THR ALA VAL TYR TYR CYS ALA VAL THR TYR ARG GLY PRO SEQRES 9 B 130 TRP PHE ASN ARG ASP PRO HIS TYR TYR TRP GLY GLN GLY SEQRES 10 B 130 THR GLN VAL THR VAL SER SER HIS HIS HIS HIS HIS HIS HET FE A 401 1 HET FE A 402 1 HET D12 A 403 12 HETNAM FE FE (III) ION HETNAM D12 DODECANE FORMUL 3 FE 2(FE 3+) FORMUL 5 D12 C12 H26 HELIX 1 AA1 ARG A 26 TRP A 29 5 4 HELIX 2 AA2 THR A 30 TRP A 35 1 6 HELIX 3 AA3 VAL A 37 THR A 49 1 13 HELIX 4 AA4 GLY A 52 TRP A 56 5 5 HELIX 5 AA5 LEU A 57 LEU A 64 1 8 HELIX 6 AA6 LEU A 64 GLY A 74 1 11 HELIX 7 AA7 VAL A 84 GLU A 89 1 6 HELIX 8 AA8 ARG A 92 LEU A 100 1 9 HELIX 9 AA9 LEU A 100 HIS A 119 1 20 HELIX 10 AB1 TYR A 120 MET A 123 5 4 HELIX 11 AB2 THR A 124 HIS A 147 1 24 HELIX 12 AB3 GLU A 148 GLY A 150 5 3 HELIX 13 AB4 THR A 154 VAL A 167 1 14 HELIX 14 AB5 HIS A 172 VAL A 185 1 14 HELIX 15 AB6 SER A 199 ARG A 225 1 27 HELIX 16 AB7 PRO A 240 GLY A 256 1 17 HELIX 17 AB8 ILE A 259 HIS A 282 1 24 HELIX 18 AB9 HIS A 308 LEU A 315 1 8 HELIX 19 AC1 ARG A 320 HIS A 327 1 8 HELIX 20 AC2 GLY A 347 TYR A 356 1 10 HELIX 21 AC3 PHE A 357 GLY A 374 1 18 HELIX 22 AC4 GLN A 386 LYS A 393 1 8 SHEET 1 AA1 4 LEU B 6 SER B 9 0 SHEET 2 AA1 4 SER B 19 ALA B 26 -1 O SER B 23 N SER B 9 SHEET 3 AA1 4 TYR B 81 ASN B 85 -1 O LEU B 82 N LEU B 22 SHEET 4 AA1 4 THR B 70 SER B 72 -1 N SER B 72 O TYR B 81 SHEET 1 AA2 4 GLU B 48 SER B 54 0 SHEET 2 AA2 4 THR B 35 GLN B 41 -1 N TRP B 38 O VAL B 50 SHEET 3 AA2 4 VAL B 94 VAL B 99 -1 O ALA B 98 N GLY B 37 SHEET 4 AA2 4 GLN B 116 GLY B 117 -1 O GLY B 117 N TYR B 95 SSBOND 1 CYS B 24 CYS B 97 1555 1555 2.04 LINK NE2 HIS A 147 FE FE A 402 1555 1555 2.41 LINK NE2 HIS A 151 FE FE A 402 1555 1555 2.26 LINK NE2 HIS A 177 FE FE A 402 1555 1555 2.25 LINK NE2 HIS A 181 FE FE A 401 1555 1555 2.33 LINK NE2 HIS A 182 FE FE A 402 1555 1555 2.29 LINK NE2 HIS A 282 FE FE A 401 1555 1555 2.16 LINK NE2 HIS A 321 FE FE A 401 1555 1555 2.31 LINK NE2 HIS A 324 FE FE A 402 1555 1555 2.23 LINK NE2 HIS A 325 FE FE A 401 1555 1555 2.15 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