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- EMDB-26460: cryo-EM structure of the ADP state wild type myosin-15-F-actin co... -
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Open data
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Basic information
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Title | cryo-EM structure of the ADP state wild type myosin-15-F-actin complex (symmetry expansion and re-centering) | ||||||||||||
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Function / homology | ![]() actin-based cell projection / Striated Muscle Contraction / stereocilium bundle / stereocilium / vesicle transport along actin filament / ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() Similarity search - Function | ||||||||||||
Biological species | ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() | ||||||||||||
Method | ![]() ![]() | ||||||||||||
![]() | Gong R / Reynolds MJ / Alushin GM | ||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural basis for tunable control of actin dynamics by myosin-15 in mechanosensory stereocilia. Authors: Rui Gong / Fangfang Jiang / Zane G Moreland / Matthew J Reynolds / Santiago Espinosa de Los Reyes / Pinar Gurel / Arik Shams / James B Heidings / Michael R Bowl / Jonathan E Bird / Gregory M Alushin / ![]() ![]() Abstract: The motor protein myosin-15 is necessary for the development and maintenance of mechanosensory stereocilia, and mutations in myosin-15 cause hereditary deafness. In addition to transporting actin ...The motor protein myosin-15 is necessary for the development and maintenance of mechanosensory stereocilia, and mutations in myosin-15 cause hereditary deafness. In addition to transporting actin regulatory machinery to stereocilia tips, myosin-15 directly nucleates actin filament ("F-actin") assembly, which is disrupted by a progressive hearing loss mutation (p.D1647G, ""). Here, we present cryo-electron microscopy structures of myosin-15 bound to F-actin, providing a framework for interpreting the impacts of deafness mutations on motor activity and actin nucleation. Rigor myosin-15 evokes conformational changes in F-actin yet maintains flexibility in actin's D-loop, which mediates inter-subunit contacts, while the mutant locks the D-loop in a single conformation. Adenosine diphosphate-bound myosin-15 also locks the D-loop, which correspondingly blunts actin-polymerization stimulation. We propose myosin-15 enhances polymerization by bridging actin protomers, regulating nucleation efficiency by modulating actin's structural plasticity in a myosin nucleotide state-dependent manner. This tunable regulation of actin polymerization could be harnessed to precisely control stereocilium height. | ||||||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 283.1 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 23.6 KB 23.6 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 18.1 KB | Display | ![]() |
Images | ![]() | 56.3 KB | ||
Masks | ![]() | 512 MB | ![]() | |
Others | ![]() ![]() | 409.8 MB 409.8 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7uduMC ![]() 7r8vC ![]() 7r91C ![]() 7rb8C ![]() 7rb9C ![]() 7udtC C: citing same article ( M: atomic model generated by this map |
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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: 1.03 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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Density Histograms |
-Half map: #2
File | emd_26460_half_map_1.map | ||||||||||||
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Density Histograms |
-Half map: #1
File | emd_26460_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
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Sample components
-Entire : the rigor state wild type myosin-15-F-actin complex
Entire | Name: the rigor state wild type myosin-15-F-actin complex |
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Components |
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-Supramolecule #1: the rigor state wild type myosin-15-F-actin complex
Supramolecule | Name: the rigor state wild type myosin-15-F-actin complex / type: complex / Chimera: Yes / ID: 1 / Parent: 0 / Macromolecule list: #1, #3 |
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Molecular weight | Experimental: 5.4 kDa/nm |
-Supramolecule #2: Actin
Supramolecule | Name: Actin / type: complex / Chimera: Yes / ID: 2 / Parent: 1 / Macromolecule list: #1 |
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Source (natural) | Organism: ![]() ![]() ![]() |
-Supramolecule #3: Isoform 3 of Unconventional myosin-XV
Supramolecule | Name: Isoform 3 of Unconventional myosin-XV / type: complex / Chimera: Yes / ID: 3 / Parent: 1 / Macromolecule list: #2 |
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Source (natural) | Organism: ![]() ![]() ![]() |
Recombinant expression | Organism: Insect cell expression vector pTIE1 (others) |
-Supramolecule #4: chicken muscle regulatory light chain
Supramolecule | Name: chicken muscle regulatory light chain / type: complex / Chimera: Yes / ID: 4 / Parent: 1 / Macromolecule list: #3 |
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Source (natural) | Organism: ![]() ![]() ![]() |
Recombinant expression | Organism: Insect cell expression vector pTIE1 (others) |
-Macromolecule #1: Actin, alpha skeletal muscle
Macromolecule | Name: Actin, alpha skeletal muscle / type: protein_or_peptide / ID: 1 / Number of copies: 3 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() ![]() |
Molecular weight | Theoretical: 41.63143 KDa |
Sequence | String: DETTALVCDN GSGLVKAGFA GDDAPRAVFP SIVGRPRHQG VMVGMGQKDS YVGDEAQSKR GILTLKYPIE (HIC)GIITN WDD MEKIWHHTFY NELRVAPEEH PTLLTEAPLN PKANREKMTQ IMFETFNVPA MYVAIQAVLS LYASGRTTGI VLDSGDG VT HNVPIYEGYA ...String: DETTALVCDN GSGLVKAGFA GDDAPRAVFP SIVGRPRHQG VMVGMGQKDS YVGDEAQSKR GILTLKYPIE (HIC)GIITN WDD MEKIWHHTFY NELRVAPEEH PTLLTEAPLN PKANREKMTQ IMFETFNVPA MYVAIQAVLS LYASGRTTGI VLDSGDG VT HNVPIYEGYA LPHAIMRLDL AGRDLTDYLM KILTERGYSF VTTAEREIVR DIKEKLCYVA LDFENEMATA ASSSSLEK S YELPDGQVIT IGNERFRCPE TLFQPSFIGM ESAGIHETTY NSIMKCDIDI RKDLYANNVM SGGTTMYPGI ADRMQKEIT ALAPSTMKIK IIAPPERKYS VWIGGSILAS LSTFQQMWIT KQEYDEAGPS IVHRKCF |
-Macromolecule #2: Unconventional myosin-XV
Macromolecule | Name: Unconventional myosin-XV / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() ![]() |
Molecular weight | Theoretical: 82.393773 KDa |
Recombinant expression | Organism: Insect cell expression vector pTIE1 (others) |
Sequence | String: EDGVEDMTQL EDLQETTVLA NLKTRFERNL IYTYIGSILV SVNPYRMFAI YGPEQVQQYS GRALGENPPH LFAIANLAFA KMLDAKQNQ CVIISGESGS GKTEATKLIL RCLAAMNQRR DVMQQIKILE ATPLLEAFGN AKTVRNDNSS RFGKFVEIFL E GGVICGAI ...String: EDGVEDMTQL EDLQETTVLA NLKTRFERNL IYTYIGSILV SVNPYRMFAI YGPEQVQQYS GRALGENPPH LFAIANLAFA KMLDAKQNQ CVIISGESGS GKTEATKLIL RCLAAMNQRR DVMQQIKILE ATPLLEAFGN AKTVRNDNSS RFGKFVEIFL E GGVICGAI TSQYLLEKSR IVFQAKNERN YHIFYELLAG LPAQLRQAFS LQEAETYYYL NQGGNCEIAG KSDADDFRRL LA AMEVLGF TSEDQDSIFR ILASILHLGN VYFEKHETDA QEVASVVSAR EIQAVAELLQ VSPEGLQKAI TFKVTETIRE KIF TPLTVE SAVDARDAIA KVLYALLFGW LITRVNALVS PKQDTLSIAI LDIYGFEDLS FNSFEQLCIN YANENLQYLF NKIV FQEEQ EEYIREQMDW REIAFADNQP CINLISLKPY GILRILDDQC CFPQATDHTF LQKCHYHHGA NPLYSKPKMP LPEFT IKHY AGKVTYQVHK FLDKNHDQVR QDVLDLFVHS RTRVVAHLFS SHAAQTAPPR LGKSSSITRL YKAHTVAAKF QQSLLD LVE KMERCNPLFV RCLKPNHKKE PGLFEPDVMM AQLRYSGVLE TVRIRKEGFP VRLPFQVFID RYRCLVALKL NVPADGD MC VSLLSRLCTV TPDMYRVGIS KLFLKEHLHQ LLESMRERVQ NRAALTLQRY LRGFFIQRHF RSLRRKIILL QSRARGF |
-Macromolecule #3: regulatory light chain
Macromolecule | Name: regulatory light chain / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() ![]() |
Molecular weight | Theoretical: 16.936826 KDa |
Recombinant expression | Organism: Insect cell expression vector pTIE1 (others) |
Sequence | String: VFAMFDQSQI QEFKEAFNMI DQNRDGFIDK EDLHDMLASL GKNPTDEYLD AMMNEAPGPI NFTMFLTMFG EKLNGTDPED VIRNAFACF DEEATGFIQE DYLRELLTTM GDRFTDEEVD ELYREAPIDK KGNFNYIEFT RILKHGA |
-Macromolecule #4: ADENOSINE-5'-DIPHOSPHATE
Macromolecule | Name: ADENOSINE-5'-DIPHOSPHATE / type: ligand / ID: 4 / Number of copies: 4 / Formula: ADP |
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Molecular weight | Theoretical: 427.201 Da |
Chemical component information | ![]() ChemComp-ADP: |
-Macromolecule #5: MAGNESIUM ION
Macromolecule | Name: MAGNESIUM ION / type: ligand / ID: 5 / Number of copies: 4 / Formula: MG |
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Molecular weight | Theoretical: 24.305 Da |
-Experimental details
-Structure determination
Method | ![]() |
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Aggregation state | filament |
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Sample preparation
Buffer | pH: 7 Details: 10 mM imidazole pH 7.0, 50 mM KCl, 1 mM MgCl2, 1 mM EGTA, 0.5 mM DTT, 0.01% NaN3 |
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Grid | Model: C-flat-1.2/1.3 / Support film - Material: CARBON / Support film - topology: HOLEY |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 293 K / Instrument: LEICA EM GP |
Details | the ADP state wild type myosin-15 bound to F-actin |
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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 | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Digitization - Frames/image: 1-40 / Number grids imaged: 1 / Average exposure time: 10.0 sec. / Average electron dose: 60.0 e/Å2 |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |