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Open data
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Basic information
Entry | ![]() | ||||||||||||
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Title | S. cerevisiae Pex1/Pex6 with 1 mM ATP | ||||||||||||
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![]() | Peroxisome AAA-ATPase unfoldase / MOTOR PROTEIN | ||||||||||||
Function / homology | ![]() protein import into peroxisome matrix, receptor recycling / protein import into peroxisome matrix / protein transporter activity / peroxisomal membrane / ATPase complex / protein unfolding / Hydrolases; Acting on acid anhydrides; Acting on acid anhydrides to facilitate cellular and subcellular movement / peroxisome / ATP hydrolysis activity / ATP binding / cytosol Similarity search - Function | ||||||||||||
Biological species | ![]() ![]() | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.89 Å | ||||||||||||
![]() | Gardner BM / Richardson CD / Martin A / Lander GC / Chowdhury S | ||||||||||||
Funding support | ![]()
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![]() | ![]() Title: The N1 domain of the peroxisomal AAA-ATPase Pex6 is required for Pex15 binding and proper assembly with Pex1. Authors: Bashir A Ali / Ryan M Judy / Saikat Chowdhury / Nicole K Jacobsen / Dominic T Castanzo / Kaili L Carr / Chris D Richardson / Gabriel C Lander / Andreas Martin / Brooke M Gardner / ![]() ![]() Abstract: The heterohexameric ATPases associated with diverse cellular activities (AAA)-ATPase Pex1/Pex6 is essential for the formation and maintenance of peroxisomes. Pex1/Pex6, similar to other AAA-ATPases, ...The heterohexameric ATPases associated with diverse cellular activities (AAA)-ATPase Pex1/Pex6 is essential for the formation and maintenance of peroxisomes. Pex1/Pex6, similar to other AAA-ATPases, uses the energy from ATP hydrolysis to mechanically thread substrate proteins through its central pore, thereby unfolding them. In related AAA-ATPase motors, substrates are recruited through binding to the motor's N-terminal domains or N terminally bound cofactors. Here, we use structural and biochemical techniques to characterize the function of the N1 domain in Pex6 from budding yeast, Saccharomyces cerevisiae. We found that although Pex1/ΔN1-Pex6 is an active ATPase in vitro, it does not support Pex1/Pex6 function at the peroxisome in vivo. An X-ray crystal structure of the isolated Pex6 N1 domain shows that the Pex6 N1 domain shares the same fold as the N-terminal domains of PEX1, CDC48, and NSF, despite poor sequence conservation. Integrating this structure with a cryo-EM reconstruction of Pex1/Pex6, AlphaFold2 predictions, and biochemical assays shows that Pex6 N1 mediates binding to both the peroxisomal membrane tether Pex15 and an extended loop from the D2 ATPase domain of Pex1 that influences Pex1/Pex6 heterohexamer stability. Given the direct interactions with both Pex15 and the D2 ATPase domains, the Pex6 N1 domain is poised to coordinate binding of cofactors and substrates with Pex1/Pex6 ATPase activity. | ||||||||||||
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 41.9 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 20.7 KB 20.7 KB | Display Display | ![]() |
Images | ![]() | 54 KB | ||
Filedesc metadata | ![]() | 7.3 KB | ||
Others | ![]() ![]() | 77.8 MB 77.8 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8u0vMC ![]() 8u0xC 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: 1.15 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: #1
File | emd_41788_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_41788_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Pex1/Pex6 AAA-ATPase
Entire | Name: Pex1/Pex6 AAA-ATPase |
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Components |
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-Supramolecule #1: Pex1/Pex6 AAA-ATPase
Supramolecule | Name: Pex1/Pex6 AAA-ATPase / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2 |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 637 KDa |
-Macromolecule #1: Peroxisomal ATPase PEX1
Macromolecule | Name: Peroxisomal ATPase PEX1 / type: protein_or_peptide / ID: 1 / Number of copies: 3 / Enantiomer: LEVO EC number: Hydrolases; Acting on acid anhydrides; Acting on acid anhydrides to facilitate cellular and subcellular movement |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 118.653977 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MTTTKRLKFE NLRIQFSNAI VGNFLRLPHS IINVLESTNY AIQEFGIAVH SHNSDIPIVH LGWDGHDSGS SENVVLINPV LATVYDLNQ KSPLVDLYIQ RYDHTHLATE VYVTPETSDD WEIIDANAMR FQNGEILHQT RIVTPGETLI CYLEGIVTKF K IDRVEPSM ...String: MTTTKRLKFE NLRIQFSNAI VGNFLRLPHS IINVLESTNY AIQEFGIAVH SHNSDIPIVH LGWDGHDSGS SENVVLINPV LATVYDLNQ KSPLVDLYIQ RYDHTHLATE VYVTPETSDD WEIIDANAMR FQNGEILHQT RIVTPGETLI CYLEGIVTKF K IDRVEPSM KSARITDGSL VVVAPKVNKT RLVKAEYGHS NKTILKNGAI QLLKKVILRS TVCKMDFPKD NLFVVYISDG AQ LPSQKGY ASIVKCSLRQ SKKSDSDNKS VGIPSKKIGV FIKCDSQIPE NHIALSSHLW DAFFTHPMNG AKIKLEFLQM NQA NIISGR NATVNIKYFG KDVPTKSGDQ YSKLLGGSLL TNNLILPTEQ IIIEIKKGES EQQLCNLNEI SNESVQWKVT QMGK EEVKD IIERHLPKHY HVKETGEVSR TSKDEDDFIT VNSIKKEMVN YLTSPIIATP AIILDGKQGI GKTRLLKELI NEVEK DHHI FVKYADCETL HETSNLDKTQ KLIMEWCSFC YWYGPSLIVL DNVEALFGKP QANDGDPSNN GQWDNASKLL NFFINQ VTK IFNKDNKRIR VLFSGKQKTQ INPLLFDKHF VSETWSLRAP DKHARAKLLE YFFSKNQIMK LNRDLQFSDL SLETEGF SP LDLEIFTEKI FYDLQLERDC DNVVTRELFS KSLSAFTPSA LRGVKLTKET NIKWGDIGAL ANAKDVLLET LEWPTKYE P IFVNCPLRLR SGILLYGYPG CGKTLLASAV AQQCGLNFIS VKGPEILNKF IGASEQNIRE LFERAQSVKP CILFFDEFD SIAPKRGHDS TGVTDRVVNQ LLTQMDGAEG LDGVYILAAT SRPDLIDSAL LRPGRLDKSV ICNIPTESER LDILQAIVNS KDKDTGQKK FALEKNADLK LIAEKTAGFS GADLQGLCYN AYLKSVHRWL SAADQSEVVP GNDNIEYFSI NEHGRREENR L RLKTLLQQ DVVHETKTST SAASELTAVV TINDLLEACQ ETKPSISTSE LVKLRGIYDR FQKDRNGEMP NGENSIDIGS RL SLMGSSD YKDDDDK UniProtKB: Peroxisomal ATPase PEX1 |
-Macromolecule #2: Peroxisomal ATPase PEX6
Macromolecule | Name: Peroxisomal ATPase PEX6 / type: protein_or_peptide / ID: 2 / Number of copies: 3 / Enantiomer: LEVO EC number: Hydrolases; Acting on acid anhydrides; Acting on acid anhydrides to facilitate cellular and subcellular movement |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 117.306758 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MGSSHHHHHH SQDPMKASLT FSLSGIYAPC SISRDIYLEY GDKKAECLYG TIRLPQYGPG CTPGKIVHCV LDDSLPFCSI VVPSKLFGF MPTQPTMDFC YFEPILDNVV PVLDSVTFLI NEQLYSKLMD LPQEMQQIQF LHYKYNINSM ETVVHSRDIL T SGLCQILN ...String: MGSSHHHHHH SQDPMKASLT FSLSGIYAPC SISRDIYLEY GDKKAECLYG TIRLPQYGPG CTPGKIVHCV LDDSLPFCSI VVPSKLFGF MPTQPTMDFC YFEPILDNVV PVLDSVTFLI NEQLYSKLMD LPQEMQQIQF LHYKYNINSM ETVVHSRDIL T SGLCQILN CSPFPQGLVD FTETQLILVN DTEQKLSALK YANEDEEYAL PKIGTNSALS IDLESLPCTI SRDLLRPAPH IN DDNSIYA FTDAETLLRL DVTSGSFITV SNMGCVRLVK LFVLLLPNGF KKRTIYAPPK IIASFPDCSV VTISKSNIGH TDI PIANQV FISRVGGWLQ SQKCFQNIIL TTLKKFFSES KRILCQNDLI PIAFDSSMAD LNIAEENDES DDEDELGQYY KNDS LVWFF VTSAELDCFS KDNSHFIIDP NRTKLITTNI TNRRPLPLSR SNLQRYYGFA ETFYYDLHIF PYVRQLVNIL ETSFN CSQR GITLNASVLL HSTTNNVGKA TMVRFASKYL GIHLLEIDCL SLTSNSRQLD STSKIIGYIR AKCENVLPYA SPAVIF LAH LDSILLDVNA NQDPEAIKLQ KSINFEMSKL LDDFTFKFPG TTFVGSVNNI DNVPSSFRSH MRFEILVPVP SEAQRLR IF QWYLSSHELN RDVQQKVPVS YMDNISFSSL SSYSAGLTPL DIKSIVETAR MTATARFYQE SKKCGWLPQS ILITQEDL S KATSKARNEF SVSIGAPQIP NVTWDDIGGI DFVKGEILDT IDMPLKHPEL FTSGMKKRSG ILFYGPPGTG KTLMAKAIA TNFSLNFFSV KGPELLNMYI GESEANVRRV FQKAREAKPC VIFFDEIDSV APKRGNQGDS GGVMDRIVSQ LLAELDGMST DADGVFVIG ATNRPDLLDE ALLRPGRFDK LLYLGIPDTD TKQLNILEAL TRKFVLDNDV KLIELAKLCP FNYTGADFYA L CSDAMLNA MSRIARMVEK KVSQHNELTG ENISTRRWFD KIATKEDTKV VVKMEDFLKA QEQLTPSVSR AELNHYEAVR AN FEGA UniProtKB: Peroxisomal ATPase PEX6 |
-Macromolecule #3: ADENOSINE-5'-TRIPHOSPHATE
Macromolecule | Name: ADENOSINE-5'-TRIPHOSPHATE / type: ligand / ID: 3 / Number of copies: 10 / Formula: ATP |
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Molecular weight | Theoretical: 507.181 Da |
Chemical component information | ![]() ChemComp-ATP: |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 3.5 mg/mL | ||||||||||||||||||||||||
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Buffer | pH: 7.6 Component:
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Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Temperature | Min: 83.0 K / Max: 83.0 K |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average exposure time: 12.0 sec. / Average electron dose: 54.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | C2 aperture diameter: 50.0 µm / Calibrated magnification: 43478 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 1.8 µm / Nominal defocus min: 0.8 µm / Nominal magnification: 43478 |
Experimental equipment | ![]() Model: Talos Arctica / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Initial model | Chain - Source name: AlphaFold / Chain - Initial model type: in silico model |
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Output model | ![]() PDB-8u0v: |