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- EMDB-14049: Cryo-EM structure of ABC transporter STE6-2p from Pichia pastoris... -
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Open data
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Basic information
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Title | Cryo-EM structure of ABC transporter STE6-2p from Pichia pastoris in apo conformation at 3.1 A resolution | |||||||||
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![]() | ABC transporter / ABCB / MDR / membrane protein / Pichia pastoris / TRANSPORT PROTEIN | |||||||||
Function / homology | ![]() ABC-type transporter activity / ATP hydrolysis activity / ATP binding / membrane Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.2 Å | |||||||||
![]() | Schleker ESM / Reinhart C | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural and functional investigation of ABC transporter STE6-2p from reveals unexpected interaction with sterol molecules. Authors: E Sabine M Schleker / Sabine Buschmann / Hao Xie / Sonja Welsch / Hartmut Michel / Christoph Reinhart / ![]() Abstract: Adenosine triphosphate (ATP)-binding cassette (ABC) transporters are multidomain transmembrane proteins, which facilitate the transport of various substances across cell membranes using energy ...Adenosine triphosphate (ATP)-binding cassette (ABC) transporters are multidomain transmembrane proteins, which facilitate the transport of various substances across cell membranes using energy derived from ATP hydrolysis. They are important drug targets since they mediate decreased drug susceptibility during pharmacological treatments. For the methylotrophic yeast , a model organism that is a widely used host for protein expression, the role and function of its ABC transporters is unexplored. In this work, we investigated the ABC-B transporter STE6-2p. Functional investigations revealed that STE6-2p is capable of transporting rhodamines in vivo and is active in the presence of verapamil and triazoles in vitro. A phylogenetic analysis displays homology among multidrug resistance (MDR) transporters from pathogenic fungi to human ABC-B transporters. Further, we present high-resolution single-particle electron cryomicroscopy structures of an ABC transporter from in the apo conformation (3.1 Å) and in complex with verapamil and adenylyl imidodiphosphate (AMP-PNP) (3.2 Å). An unknown density between transmembrane helices 4, 5, and 6 in both structures suggests the presence of a sterol-binding site of unknown function. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 57.3 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 16.2 KB 16.2 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 11.5 KB | Display | ![]() |
Images | ![]() | 91.6 KB | ||
Masks | ![]() | 91.1 MB | ![]() | |
Filedesc metadata | ![]() | 6.4 KB | ||
Others | ![]() ![]() | 71.2 MB 71.2 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 817.9 KB | Display | ![]() |
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Full document | ![]() | 817.4 KB | Display | |
Data in XML | ![]() | 17.5 KB | Display | |
Data in CIF | ![]() | 23 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7qkrMC ![]() 7qksC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Voxel size | X=Y=Z: 0.837 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
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-Half map: #2
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Density Histograms |
-Half map: #1
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Sample components
-Entire : STE6-2p
Entire | Name: STE6-2p |
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Components |
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-Supramolecule #1: STE6-2p
Supramolecule | Name: STE6-2p / type: organelle_or_cellular_component / ID: 1 / Parent: 0 / Macromolecule list: #1 |
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Source (natural) | Organism: ![]() |
-Macromolecule #1: Plasma membrane ATP-binding cassette transporter required for the...
Macromolecule | Name: Plasma membrane ATP-binding cassette transporter required for the export of a-factor type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() Strain: ATCC 76273 / CBS 7435 / CECT 11047 / NRRL Y-11430 / Wegner 21-1 |
Molecular weight | Theoretical: 144.974266 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MVKNYDYKDD DDKGLSFFKR KKYSDKESLS SPETLEEKPN DELDPRVTEI LERQIKADSY GASLVDLYGM LQGWEYCLAV AAYICSIVA GAALPLMTLI FGDMAQQFTD YSSGLHSNNQ FVDKIDENAL YFVYLGVGLL VFNYFATLLH IVVSEIIASR V REKFIWSI ...String: MVKNYDYKDD DDKGLSFFKR KKYSDKESLS SPETLEEKPN DELDPRVTEI LERQIKADSY GASLVDLYGM LQGWEYCLAV AAYICSIVA GAALPLMTLI FGDMAQQFTD YSSGLHSNNQ FVDKIDENAL YFVYLGVGLL VFNYFATLLH IVVSEIIASR V REKFIWSI LHQNMAYLDS LGSGEITSSI TSDSQLIQQG VSEKIGLAAQ SIATVVSALT VAFVIYWKLA LVLLSVMVAL IL SSTPTIL MLMQAYTDSI ASYGKASSVA EEAFAAIKTA TAFGAHEFQL QKYDEFILES KGYGKKKAIS LALMMGSIWF IVF ATYALA FWQGSRFMVS DNSGIGKILT ACMAMLFGSL IIGNATISLK FVMVGLSAAS KLFAMINREP YFDSASDAGE KINE FDGSI SFRNVTTRYP SRPDITVLSD FTLDIKPGQT IALVGESGSG KSTVIALLER FYEYLDGEIL LDGVDLKSLN IKWVR QQMA LVQQEPVLFA ASIYENVCYG LVGSKYENVT EKVKRELVEK ACKDANAWEF ISQMSNGLDT EVGERGLSLS GGQKQR IAI ARAVISEPKI LLLDEATSAL DTRSEGIVQD ALNRLSETRT TIVIAHRLST IQNADLIVVL SKGKIVETGS HKELLKK KG KYHQLVQIQN IRTKINNSGP QAPISLSNSS DLDSVSHKID RVESLIYERA AADTIDESPV KKQSIPQLFL MLLQINKG D YYLLIPCLFL ALIAGMGFPS FALLAGRVIE AFQVTGPQDF PHMRSLINKY TGFLFMIGCV LLIVYLFLTS FMVLSSESL VYKMRYRCFK QYLRQDMSFF DRPENKVGTL VTTLAKDPQD IEGLSGGTAA QLAVSVVIVV AGIILAVAVN WRLGLVCTAT VPILLGCGF FSVYLLMVFE ERILKDYQES ASYACEQVSA LKTVVSLTRE VGIYEKYSNS IKDQVKRSAR SVSRTTLLYA L IQGMNPWV FALGFWYGSR LLLEGRATNR EFFTVLMAIL FGCQSAGEFF SYAPGMGKAK QAAINIRQVL DTRPKSIDIE SE DGLKIDR LNLKGGIELR DVTFRYPTRP EVPVLTDLNL IIKPGQYVGL VGASGCGKST TVGLIERFYD PESGQVLLDG VDI RDLHLR TYREVLALVQ QEPVLFSGSI RDNIMVGSIS DGADDGSEED MIKACKDANI YDFISSLPEG FDTLCGNKGT MLSG GQKQR VAIARALIRN PRVLLLDEAT SALDSESEMV VQDAIDKASK GRTTITIAHR LSTVQNCDVI YVFDAGRIVE SGKHD ELLQ LRGKYYDLVQ LQGLNAHHHH HHHHHH UniProtKB: Plasma membrane ATP-binding cassette transporter required for the export of a-factor |
-Macromolecule #2: Dexverapamil
Macromolecule | Name: Dexverapamil / type: ligand / ID: 2 / Number of copies: 1 / Formula: I6H |
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Molecular weight | Theoretical: 454.602 Da |
Chemical component information | ![]() ChemComp-I6H: |
-Macromolecule #3: PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER
Macromolecule | Name: PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER / type: ligand / ID: 3 / Number of copies: 1 / Formula: ANP |
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Molecular weight | Theoretical: 506.196 Da |
Chemical component information | ![]() ChemComp-ANP: |
-Macromolecule #4: MAGNESIUM ION
Macromolecule | Name: MAGNESIUM ION / type: ligand / ID: 4 / Number of copies: 1 / Formula: MG |
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Molecular weight | Theoretical: 24.305 Da |
-Macromolecule #5: (3beta,14beta,17beta,25R)-3-[4-methoxy-3-(methoxymethyl)butoxy]sp...
Macromolecule | Name: (3beta,14beta,17beta,25R)-3-[4-methoxy-3-(methoxymethyl)butoxy]spirost-5-en type: ligand / ID: 5 / Number of copies: 1 / Formula: 9Z9 |
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Molecular weight | Theoretical: 544.805 Da |
Chemical component information | ![]() ChemComp-9Z9: |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 2.5 mg/mL |
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Buffer | pH: 8 |
Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 40.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: OTHER / Imaging mode: BRIGHT FIELD / Nominal defocus max: 0.0021000000000000003 µm / Nominal defocus min: 0.0011 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |