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- EMDB-42513: Structural and biochemical investigations of a HEAT-repeat protei... -
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Open data
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Basic information
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Title | Structural and biochemical investigations of a HEAT-repeat protein involved in the cytosolic iron-sulfur cluster assembly pathway | |||||||||||||||
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![]() | cytosolic iron-sulfur cluster pathway / ![]() ![]() ![]() | |||||||||||||||
Biological species | ![]() ![]() ![]() | |||||||||||||||
Method | ![]() ![]() | |||||||||||||||
![]() | Vasquez S / Drennan C | |||||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural and biochemical investigations of a HEAT-repeat protein involved in the cytosolic iron-sulfur cluster assembly pathway. Authors: Sheena Vasquez / Melissa D Marquez / Edward J Brignole / Amanda Vo / Sunnie Kong / Christopher Park / Deborah L Perlstein / Catherine L Drennan / ![]() Abstract: Iron-sulfur clusters are essential for life and defects in their biosynthesis lead to human diseases. The mechanism of cluster assembly and delivery to cytosolic and nuclear client proteins via the ...Iron-sulfur clusters are essential for life and defects in their biosynthesis lead to human diseases. The mechanism of cluster assembly and delivery to cytosolic and nuclear client proteins via the cytosolic iron-sulfur cluster assembly (CIA) pathway is not well understood. Here we report cryo-EM structures of the HEAT-repeat protein Met18 from Saccharomyces cerevisiae, a key component of the CIA targeting complex (CTC) that identifies cytosolic and nuclear client proteins and delivers a mature iron-sulfur cluster. We find that in the absence of other CTC proteins, Met18 adopts tetrameric and hexameric states. Using mass photometry and negative stain EM, we show that upon the addition of Cia2, these higher order oligomeric states of Met18 disassemble. We also use pulldown assays to identify residues of critical importance for Cia2 binding and recognition of the Leu1 client, many of which are buried when Met18 oligomerizes. Our structures show conformations of Met18 that have not been previously observed in any Met18 homolog, lending support to the idea that a highly flexible Met18 may be key to how the CTC is able to deliver iron-sulfur clusters to client proteins of various sizes and shapes, i.e. Met18 conforms to the dimensions needed. | |||||||||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 11.3 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 17.5 KB 17.5 KB | Display Display | ![]() |
Images | ![]() | 128.5 KB | ||
Filedesc metadata | ![]() | 5.7 KB | ||
Others | ![]() ![]() | 108.6 MB 108.5 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
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Voxel size | X=Y=Z: 1.059 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: #2
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_42513_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Met18/MMS19
Entire | Name: Met18/MMS19 |
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Components |
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-Supramolecule #1: Met18/MMS19
Supramolecule | Name: Met18/MMS19 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() ![]() ![]() |
Molecular weight | Theoretical: 118 kDa/nm |
-Macromolecule #1: Met18/MMS19
Macromolecule | Name: Met18/MMS19 / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() ![]() |
Recombinant expression | Organism: ![]() ![]() ![]() |
Sequence | String: AVVTFMANLN IDDSKANETA STVTDSIVHR SIKLLEVVVA LKDYFLSENE VERKKALTCL TTILAKTPKD HLSKNECSV IFQFYQSKLD DQALAKEVLE GFAALAPMKY VSINEIAQLL RLLLDNYQQG QHLASTRLWP F KILRKIFD RFFVNGSSTE QVKRINDLFI ...String: AVVTFMANLN IDDSKANETA STVTDSIVHR SIKLLEVVVA LKDYFLSENE VERKKALTCL TTILAKTPKD HLSKNECSV IFQFYQSKLD DQALAKEVLE GFAALAPMKY VSINEIAQLL RLLLDNYQQG QHLASTRLWP F KILRKIFD RFFVNGSSTE QVKRINDLFI ETFLHVANGE KDPRNLLLSF ALNKSITSSL QNVENAKEDL FD VLFCYFA ALKTALRSAI TATPLFAEDA YSNLLDKLTA SSPVVKNDTL LTLLECVRKF GGSSILENWT LLW NALKFE IMQNYTNYDA CLKIINLMAL QLYNFDKVSF EKFFTHVLDE LKPNFKYEKD LKQTCQILSA IGSG NVEIF NKVISSTFPL FLINTSEVAK LKLLIMNFSF FVDSYIDLFG RTSKESLGTP VPNNKMAEYK DEIIM ILSM ALTRSSKAEV TIRTLSVIQF TKMIKMKGFL TPEEVSLIIQ YFTEEILTDN NKNIYYACLE GLKTIS EIY EDLVFEISLK KLLDLLPDCF EEKIRVNDEE NIHIETILKI ILDFTTSRHI LVKESITFLA TKLNRVA KI SKSREYCFLL ISTIYSLFNN NNQNENVLNE EDALALKNAI EPKLFEIITQ ESAIVSDNYN LTLLSNVL F FTNLKIPQAA HQEELDRYNE LFISEGKIRI LDTPNVLAIS YAKILSALNK NCQFPQKFTV LFGTVQLLK KHAPRMTETE KLGYLELLLV LSNKFVSEKD VIGLFDWKDL SVINLEVMVW LTKGLIMQNS LESSEIAKKF IDLLSNEEI GSLVSKLFEV FVMDISSLKK FKGISWNNNV KILYKQKFFG DIFQTLVSNY KNTVDMTIKC N YLTALSLV LKHTPSQSVG PFINDLFPLL LQALDMPDPE VRVSALETLK DTTDKHHTLI TEHVSTIVPL LL SLSLPHK YNSVSVRLIA LQLLEMITTV VPLNYCLSYQ DDVLSALIPV LSDKKRIIRK QCVDTRQVYY ELG QI |
-Experimental details
-Structure determination
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Aggregation state | particle |
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Sample preparation
Concentration | 1.2 mg/mL | ||||||||||
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Buffer | pH: 7.5 Component:
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Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 283.15 K / Instrument: FEI VITROBOT MARK I |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD![]() |
Sample stage | Cooling holder cryogen: NITROGEN |
Image recording | Film or detector model: GATAN K2 QUANTUM (4k x 4k) / Average electron dose: 49.59 e/Å2 |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Particle selection | Number selected: 379779 |
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Startup model | Type of model: NONE |
Initial angle assignment | Type: NOT APPLICABLE |
Final 3D classification | Number classes: 3 / Avg.num./class: 50000 / Software - Name: RELION |
Final angle assignment | Type: NOT APPLICABLE |
Final reconstruction | Applied symmetry - Point group: D3 (2x3 fold dihedral![]() |
-Atomic model buiding 1
Initial model | PDB ID: Chain - Source name: PDB / Chain - Initial model type: experimental model |
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Refinement | Protocol: RIGID BODY FIT |