EMDB-55043: CM1-activated gTuRC in complex with nascent alpha-E254D mutant mi...

Basic information

Entry

Database: EMDB / ID: EMD-55043

• Title: CM1-activated gTuRC in complex with nascent alpha-E254D mutant microtubules
• Map data: Consensus map

Sample

• Complex: gamma-tubulin ring complex
  • Protein or peptide: x 12 types
• Ligand: x 1 types

• Keywords: Microtubule / cytoskeleton / g-tubulin ring complex / tubulin / STRUCTURAL PROTEIN / CDK5RAP2 / CM1 / nucleation

Function / homology

Function:

basal body patch / protein localization to astral microtubule / protein localization to mitotic spindle / cortical microtubule cytoskeleton / mitotic spindle astral microtubule end / microtubule nucleation by interphase microtubule organizing center / tight junction assembly / netrin receptor binding / gamma-tubulin complex localization / microtubule nucleator activity / protein localization to bicellular tight junction / negative regulation of centriole replication / microtubule organizing center organization / gamma-tubulin complex / regulation of mitotic cell cycle spindle assembly checkpoint / protein localization to microtubule / Post-chaperonin tubulin folding pathway / dorsal root ganglion development / cytoskeleton-dependent intracellular transport / Cargo trafficking to the periciliary membrane / SUMO is conjugated to E1 (UBA2:SAE1) / SUMOylation of nuclear envelope proteins / polar microtubule / SUMO is transferred from E1 to E2 (UBE2I, UBC9) / Carboxyterminal post-translational modifications of tubulin / SUMO is proteolytically processed / SUMOylation of SUMOylation proteins / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / SUMOylation of transcription factors / gamma-tubulin ring complex / interphase microtubule organizing center / SUMOylation of transcription cofactors / profilin binding / Postmitotic nuclear pore complex (NPC) reformation / cell projection membrane / mitotic spindle microtubule / septin ring / Sealing of the nuclear envelope (NE) by ESCRT-III / SUMOylation of DNA damage response and repair proteins / Intraflagellar transport / Transcriptional and post-translational regulation of MITF-M expression and activity / SUMOylation of RNA binding proteins / SUMOylation of DNA replication proteins / attachment of mitotic spindle microtubules to kinetochore / regulation of transepithelial transport / microtubule plus-end binding / microtubule plus-end / morphogenesis of a polarized epithelium / Formation of annular gap junctions / Formation of the dystrophin-glycoprotein complex (DGC) / Formation of tubulin folding intermediates by CCT/TriC / structural constituent of postsynaptic actin cytoskeleton / Gap junction degradation / GBP-mediated host defense / SUMOylation of chromatin organization proteins / gamma-tubulin binding / Cell-extracellular matrix interactions / microtubule bundle formation / Gap junction assembly / dense body / regulation of stress fiber assembly / single fertilization / Regulation of CDH1 Function / Kinesins / meiotic spindle organization / non-motile cilium / Adherens junctions interactions / Prefoldin mediated transfer of substrate to CCT/TriC / microtubule nucleation / Assembly and cell surface presentation of NMDA receptors / COPI-independent Golgi-to-ER retrograde traffic / Sensory processing of sound by outer hair cells of the cochlea / centrosome cycle / regulation of neuron differentiation / negative regulation of neuron differentiation / regulation of focal adhesion assembly / sperm principal piece / Sensory processing of sound by inner hair cells of the cochlea / COPI-dependent Golgi-to-ER retrograde traffic / maintenance of blood-brain barrier / Interaction between L1 and Ankyrins / apical junction complex / positive regulation of wound healing / mitotic spindle pole / intercellular bridge / spindle midzone / negative regulation of microtubule polymerization / sperm end piece / centriole replication / NuA4 histone acetyltransferase complex / pericentriolar material / microtubule organizing center / cell leading edge / filamentous actin / Recycling pathway of L1 / mitotic sister chromatid segregation / microtubule polymerization / myofibril / ubiquitin-like protein ligase binding / RHOH GTPase cycle

Domain/homology:

CDK5 regulatory subunit-associated protein 2 / : / : / CDK5 regulatory subunit-associated protein 2-like, coiled coil / Myomegalin domain / MOZART2 family / Mitotic-spindle organizing gamma-tubulin ring associated / Centrosomin, N-terminal motif 1 / Centrosomin N-terminal motif 1 / Mitotic-spindle organizing protein 1 / Mitotic-spindle organizing gamma-tubulin ring associated / EB1, C-terminal / Microtubule-associated protein RP/EB / EB1, C-terminal domain superfamily / EB1-C terminal (EB1-C) domain profile. / : / EB1-like C-terminal motif / Gamma-tubulin complex component 6, N-terminal / Gamma-tubulin complex component 6 N-terminus / Gamma tubulin / Gamma tubulin complex component, C-terminal / Gamma-tubulin complex component, C-terminal domain superfamily / Gamma tubulin complex component C-terminal / Gamma-tubulin complex component protein / Gamma tubulin complex component protein, N-terminal / Gamma tubulin complex component N-terminal / Rad60/SUMO-like domain / Ubiquitin-2 like Rad60 SUMO-like / Calponin homology (CH) domain / Calponin homology domain / CH domain superfamily / Calponin homology (CH) domain profile. / Alpha tubulin / Tubulin-beta mRNA autoregulation signal. / Beta tubulin, autoregulation binding site / Beta tubulin / Actins signature 1. / Actin, conserved site / Actins signature 2. / Actin/actin-like conserved site / Actins and actin-related proteins signature. / Tubulin / Tubulin, C-terminal / Tubulin C-terminal domain / Tubulin, conserved site / Tubulin subunits alpha, beta, and gamma signature. / Actin / Actin family / Actin / Tubulin/FtsZ family, C-terminal domain / Tubulin/FtsZ-like, C-terminal domain / Tubulin/FtsZ, C-terminal / Tubulin/FtsZ, 2-layer sandwich domain / Tubulin/FtsZ family, GTPase domain / Tubulin/FtsZ family, GTPase domain / Tubulin/FtsZ, GTPase domain / Tubulin/FtsZ, GTPase domain superfamily / ATPase, nucleotide binding domain / Ubiquitin homologues / Ubiquitin domain profile. / Ubiquitin-like domain / Ubiquitin-like domain superfamily

Component:

Tubulin gamma-1 chain / Actin, cytoplasmic 2 / Tubulin alpha-1B chain / Mitotic-spindle organizing protein 1 / Small ubiquitin-related modifier / Tubulin beta-3 chain / Microtubule-associated protein RP/EB family member 1 / Mitotic-spindle organizing protein 2A / Gamma-tubulin complex component 3 / Gamma-tubulin complex component 6 / Gamma-tubulin complex component 5 / CDK5 regulatory subunit-associated protein 2 / Gamma-tubulin complex component 2 / Gamma-tubulin complex component 4

• Biological species: Homo sapiens (human)
• Method: single particle reconstruction / cryo EM / Resolution: 3.67 Å
• Authors: Llorca O / Serna M / Lopez-Perrote A

Funding support

Spain, European Union, 2 items

Organization	Grant number	Country
Agencia Estatal de Investigacion (AEI)	MCIN/AEI/10.13039/501100011033	Spain
European Regional Development Fund	PID2023-146110NB-I00	European Union

• Citation: Journal: Nat Commun / Year: 2026; Title: Structural basis of human γTuRC closure during CM1-activated microtubule nucleation.; Authors: Marina Serna / Cláudia Brito / Silvia Speroni / Fabian Zimmermann / Andrés Lopez-Perrote / Maria Gili / Cristina Lacasa / Jens Lüders / Thomas Surrey / Oscar Llorca / ; Abstract: Microtubule nucleation by the γ-tubulin ring complex (γTuRC) is spatiotemporally regulated and in higher eukaryotes is thought to involve a transition from an inactive open to an active closed conformation that matches the microtubule geometry. However, γTuRC activators only promote a partially closed conformation, raising the question of whether complete closure is required for activation. Combining in vitro nucleation assays and cryo-EM, we find that centrosomin motif 1 (CM1), a conserved element of several γTuRC regulators, potently accelerates human γTuRC-mediated microtubule nucleation by facilitating complete closure of γTuRC as the nascent microtubule assembles. A 3.7 Å cryo-EM structure identifies the γTuRC latch and several interactions involved in conformational closure. Notably, the distinct subunits that keep γTuRC open and inactive in higher eukaryotes also participate in its closure and activation. This work provides additional insight into the logic of the human γTuRC architecture and its activation by CM1.

History

Deposition	Sep 9, 2025	-
Header (metadata) release	May 13, 2026	-
Map release	May 13, 2026	-
Update	Jun 3, 2026	-
Current status	Jun 3, 2026	Processing site: PDBe / Status: Released

Map

• File: Download / File: emd_55043.map.gz / Format: CCP4 / Size: 614.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
• Annotation: Consensus map
• Voxel size: X=Y=Z: 1.06 Å

Density

Contour Level	By AUTHOR: 0.0035
Minimum - Maximum	-0.0048037297 - 0.018327953
Average (Standard dev.)	-0.00015725355 (±0.00083585107)

• Symmetry: Space group: 1

Details

EMDB XML:

Map geometry	Axis order X Y Z Origin 0 0 0 Dimensions 544 544 544 Spacing 544 544 544
Cell	A=B=C: 576.63995 Å α=β=γ: 90.0 °

Supplemental data

Mask #1

• File: emd_55043_msk_1.map

Additional map: Focused refined map for the modules in spokes 3, 5 and 7

• File: emd_55043_additional_1.map
• Annotation: Focused refined map for the modules in spokes 3, 5 and 7

Additional map: Consensus map sharpened using deepEMhancer

• File: emd_55043_additional_2.map
• Annotation: Consensus map sharpened using deepEMhancer

Additional map: Focused refined map for the module in spoke 13

• File: emd_55043_additional_3.map
• Annotation: Focused refined map for the module in spoke 13

Additional map: Sharpened focused refined map for the modules in...

• File: emd_55043_additional_4.map
• Annotation: Sharpened focused refined map for the modules in spokes 3, 5 and 7 using deepEMhancer

Additional map: Sharpened focused refined map for the module in...

• File: emd_55043_additional_5.map
• Annotation: Sharpened focused refined map for the module in spoke 13 using deepEMhancer

Additional map: Sharpened consensus map

• File: emd_55043_additional_6.map
• Annotation: Sharpened consensus map

Additional map: Sharpened focused refined map for the gTuRC seam using deepEMhancer

• File: emd_55043_additional_7.map
• Annotation: Sharpened focused refined map for the gTuRC seam using deepEMhancer

Additional map: Focused refined map for the gTuRC seam

• File: emd_55043_additional_8.map
• Annotation: Focused refined map for the gTuRC seam

Half map: Half 1 map of the consensus refinement

• File: emd_55043_half_map_1.map
• Annotation: Half 1 map of the consensus refinement

Half map: Half 2 map of the consensus refinement

• File: emd_55043_half_map_2.map
• Annotation: Half 2 map of the consensus refinement

Sample components

Entire : gamma-tubulin ring complex

• Entire: Name: gamma-tubulin ring complex

Components

• Complex: gamma-tubulin ring complex
  • Protein or peptide: Gamma-tubulin complex component 3
  • Protein or peptide: Mitotic-spindle organizing protein 1
  • Protein or peptide: Gamma-tubulin complex component 6
  • Protein or peptide: Actin, cytoplasmic 2, N-terminally processed
  • Protein or peptide: Gamma-tubulin complex component 2
  • Protein or peptide: Ubiquitin-like protein SMT3,CDK5 regulatory subunit-associated protein 2,Microtubule-associated protein RP/EB family member 1
  • Protein or peptide: Isoform 1 of Tubulin alpha-1B chain
  • Protein or peptide: Tubulin beta-3 chain
  • Protein or peptide: Mitotic-spindle organizing protein 2A
  • Protein or peptide: Gamma-tubulin complex component 4
  • Protein or peptide: Gamma-tubulin complex component 5
  • Protein or peptide: Tubulin gamma-1 chain
• Ligand: GUANOSINE-5'-TRIPHOSPHATE

Supramolecule #1: gamma-tubulin ring complex

• Supramolecule: Name: gamma-tubulin ring complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#12
• Source (natural): Organism: Homo sapiens (human)

Macromolecule #1: Gamma-tubulin complex component 3

• Macromolecule: Name: Gamma-tubulin complex component 3 / type: protein_or_peptide / ID: 1 / Number of copies: 6 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 103.710102 KDa

Sequence

String:

MATPDQKSPN VLLQNLCCRI LGRSEADVAQ QFQYAVRVIG SNFAPTVERD EFLVAEKIKK ELIRQRREAD AALFSELHRK LHSQGVLKN KWSILYLLLS LSEDPRRQPS KVSSYATLFA QALPRDAHST PYYYARPQTL PLSYQDRSAQ SAQSSGSVGS S GISSIGLC ALSGPAPAPQ SLLPGQSNQA PGVGDCLRQQ LGSRLAWTLT ANQPSSQATT SKGVPSAVSR NMTRSRREGD TG GTMEITE AALVRDILYV FQGIDGKNIK MNNTENCYKV EGKANLSRSL RDTAVRLSEL GWLHNKIRRY TDQRSLDRSF GLV GQSFCA ALHQELREYY RLLSVLHSQL QLEDDQGVNL GLESSLTLRR LLVWTYDPKI RLKTLAALVD HCQGRKGGEL ASAV HAYTK TGDPYMRSLV QHILSLVSHP VLSFLYRWIY DGELEDTYHE FFVASDPTVK TDRLWHDKYT LRKSMIPSFM TMDQS RKVL LIGKSINFLH QVCHDQTPTT KMIAVTKSAE SPQDAADLFT DLENAFQGKI DAAYFETSKY LLDVLNKKYS LLDHMQ AMR RYLLLGQGDF IRHLMDLLKP ELVRPATTLY QHNLTGILET AVRATNAQFD SPEILRRLDV RLLEVSPGDT GWDVFSL DY HVDGPIATVF TRECMSHYLR VFNFLWRAKR MEYILTDIRK GHMCNAKLLR NMPEFSGVLH QCHILASEMV HFIHQMQY Y ITFEVLECSW DELWNKVQQA QDLDHIIAAH EVFLDTIISR CLLDSDSRAL LNQLRAVFDQ IIELQNAQDA IYRAALEEL QRRLQFEEKK KQREIEGQWG VTAAEEEEEN KRIGEFKESI PKMCSQLRIL THFYQGIVQQ FLVLLTTSSD ESLRFLSFRL DFNEHYKAR EPRLRVSLGT RGRRSSHT

UniProtKB: Gamma-tubulin complex component 3

Macromolecule #2: Mitotic-spindle organizing protein 1

• Macromolecule: Name: Mitotic-spindle organizing protein 1 / type: protein_or_peptide / ID: 2 / Number of copies: 3 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 8.485724 KDa

Sequence

String:

MASSSGAGAA AAAAAANLNA VRETMDVLLE ISRILNTGLD METLSICVRL CEQGINPEAL SSVIKELRKA TEALKAAENM TS

UniProtKB: Mitotic-spindle organizing protein 1

Macromolecule #3: Gamma-tubulin complex component 6

• Macromolecule: Name: Gamma-tubulin complex component 6 / type: protein_or_peptide / ID: 3 / Number of copies: 2 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 200.733641 KDa

Sequence

String:

MASITQLFDD LCEALLPAAK THLGQRSVNR KRAKRSLKKV AYNALFTNLF QDETQQLQPD MSKLPARNKI LMLSFDLRVG GLGPKADRL EELVEELEAA PCCPLLEVGS VLDLLVQLAG SGPPQVLPRK RDYFLNNKHV GRNVPYSGYD CDDLSVFEMD V QSLISREE CLCHSMIQET LQVMEAAPGT GLPTVGLFSF GDPCGDRFER DTRVSLFGAL VHSRTYDMDV RLGLPPVPDN AD LSGLAIK VPPSVDQWED EGFQSASNLT PDSQSEPSVT PDVDLWEAAL TYEASKRRCW ERVGCPPGHR EEPYLTEAGR DAF DKFCRL HQGELQLLAG GVLQAPQPVL VKECELVKDV LNVLIGVVSA TFSLCQPAQA FVVKRGVHVS GASPESISSL LSEV AEYGT CYTRLSHFSL QPVLDSLYSK GLVFQAFTSG LRRYLQYYRA CVLSTPPTLS LLTIGFLFKK LGRQLRYLAE LCGVG AVLP GTCGGGPRAA FPTGVKLLSY LYQEALHNCS NEHYPVLLSL LKTSCEPYTR FIHDWVYSGV FRDAYGEFMI QVNHEY LSF RDKLYWTHGY VLISKEVEDC VPVFLKHIAH DIYVCGKTIN LLKLCCPRHY LCWSDVPVPR ISVIFSLEEL KEIEKDC AV YVGRMERVAR HSSVSKEEKE LRMEIAKQEL IAHAREAASR VLSALSDRQM SERMALDARK REQFQRLKEQ FVKDQERR Q AARQEELDDD FSYARELRDR ERRLKSLEEE LERKARQALV DHYSKLSAEA ARREQKALWR IQRHRLESAR LRFLLEDEK HIQEMLKAVS EAHQPQEPPD VLLSVHPQVT SPGPEHPEGG QGCDSGSAEQ HSPAWDGWNR PGLLTPQPLK PLAVGAGGRG LQQAEGARP FSDSLSIGDF LPVGPGAEPS VQTGMVPLLE VALQTINLDL PPSAPGEAPA AASTQPSRPQ EYDFSTVLRP A VATSPAPG PLQAAECSLG SSGLQLWEDS CGKMDACGSA SRETLLPSHP PRRAALEEGS SQPTERLFGQ VSGGGLPTGD YA SEIAPTR PRWNTHGHVS DASIRVGENV SDVAPTQPRW NTHGHVSNAS ISLGESVSDV APTRPRWNIH GHVSNASIRV GEN VSDVAP TRPRWNTHGH VSNASIRVGE NVSDVAPTRP RWNTHGHVSD ASISLGESVS DMAPARPRWN THGHVSDASI SLGE SVSDM APTRPRWNTH GHVSDTSIRV GENVSDVAPI RSRCNTHGHV SDASISLGEP VSDVVSTRPR WNTHVPIPPP HMVLG ALSP EAEPNTPRPQ QSPPGHTSQS ALSLGAQSTV LDCGPRLPVE VGPSLSSPSS GCGEGSISVG ENVSDVAPTQ PWWPNT PGD SVSEELGPGR SGDTEDLSPN WPLNSQEDTA AQSSPGRGEE AEASAAEAQG GEQAYLAGLA GQYHLERYPD SYESMSE PP IAHLLRPVLP RAFAFPVDPQ VQSAADETAV QLSELLTLPV LMKRSITAPL AAHISLVNKA AVDYFFVELH LEAHYEAL R HFLLMEDGEF AQSLSDLLFE KLGAGQTPGE LLNPLVLNSV LSKALQCSLH GDTPHASNLS LALKYLPEVF APNAPDVLS CLELRYKVDW PLNIVITEGC VSKYSGVFSF LLQLKLMMWA LKDVCFHLKR TALLSHMAGS VQFRQLQLFK HEMQHFVKVI QGYIANQIL HVTWCEFRAR LATVGDLEEI QRAHAEYLHK AVFRGLLTEK AAPVMNVIHS IFSLVLKFRS QLISQAWGPP G GPRGAEHP NFALMQQSYN TFKYYSHFLF KVVTKLVNRG YQPHLEDFLL RINFNNYYQD A

UniProtKB: Gamma-tubulin complex component 6

Macromolecule #4: Actin, cytoplasmic 2, N-terminally processed

• Macromolecule: Name: Actin, cytoplasmic 2, N-terminally processed / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 41.723527 KDa

Sequence

String:

DEEVAALVVD NGSGMCKAGF AGDDAPRAVF PSIVGRPRHQ GVMVGMGQKD SYVGDEAQSK RGILTLKYPI EHGIVTNWDD MEKIWHHTF YNELRVAPEE HPVLLTEAPL NPKANREKMT QIMFETFNTP AMYVAIQAVL SLYASGRTTG IVLDSGDGVS H TVPIYEGY ALPHAILRLD LAGRDLTDYL MKILTERGYS FTTTEEREIV RDIKEKLCYV ALDFEQEMAT AASSSSLEKS YE LKDGQVI TIGNERFRCP EALFQPSFLG MEACGIHETT YNSIMKCDVD IRKDLYANTV LSGGTTMYPG IADRMQKEIT ALA PSTMKI KIIAPPERKY SVWIGGSILA SLSTFQQMWI SKQEYDESGP SIVHRKCF

UniProtKB: Actin, cytoplasmic 2

Macromolecule #5: Gamma-tubulin complex component 2

• Macromolecule: Name: Gamma-tubulin complex component 2 / type: protein_or_peptide / ID: 5 / Number of copies: 8 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 102.666953 KDa

Sequence

String:

MSEFRIHHDV NELLSLLRVH GGDGAEVYID LLQKNRTPYV TTTVSAHSAK VKIAEFSRTP EDFLKKYDEL KSKNTRNLDP LVYLLSKLT EDKETLQYLQ QNAKERAELA AAAVGSSTTS INVPAAASKI SMQELEELRK QLGSVATGST LQQSLELKRK M LRDKQNKK NSGQHLPIFP AWVYERPALI GDFLIGAGIS TDTALPIGTL PLASQESAVV EDLLYVLVGV DGRYVSAQPL AG RQSRTFL VDPNLDLSIR ELVHRILPVA ASYSAVTRFI EEKSSFEYGQ VNHALAAAMR TLVKEHLILV SQLEQLHRQG LLS LQKLWF YIQPAMRTMD ILASLATSVD KGECLGGSTL SLLHDRSFSY TGDSQAQELC LYLTKAASAP YFEVLEKWIY RGII HDPYS EFMVEEHELR KERIQEDYND KYWDQRYTIV QQQIPSFLQK MADKILSTGK YLNVVRECGH DVTCPVAKEI IYTLK ERAY VEQIEKAFNY ASKVLLDFLM EEKELVAHLR SIKRYFLMDQ GDFFVHFMDL AEEELRKPVE DITPPRLEAL LELALR MST ANTDPFKDDL KIDLMPHDLI TQLLRVLAIE TKQEKAMAHA DPTELALSGL EAFSFDYIVK WPLSLIINRK ALTRYQM LF RHMFYCKHVE RQLCSVWISN KTAKQHSLHS AQWFAGAFTL RQRMLNFVQN IQYYMMFEVM EPTWHILEKN LKSASNID D VLGHHTGFLD TCLKDCMLTN PELLKVFSKL MSVCVMFTNC MQKFTQSMKL DGELGGQTLE HSTVLGLPAG AEERARKEL ARKHLAEHAD TVQLVSGFEA TINKFDKNFS AHLLDLLARL SIYSTSDCEH GMASVISRLD FNGFYTERLE RLSAERSQKA TPQVPVLRG PPAPAPRVAV TAQ

UniProtKB: Gamma-tubulin complex component 2

Macromolecule #6: Ubiquitin-like protein SMT3,CDK5 regulatory subunit-associated pr...

• Macromolecule: Name: Ubiquitin-like protein SMT3,CDK5 regulatory subunit-associated protein 2,Microtubule-associated protein RP/EB family member 1; type: protein_or_peptide / ID: 6 / Number of copies: 9 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 26.765963 KDa
• Recombinant expression: Organism: Escherichia coli (E. coli)

Sequence

String:

MGSSHHHHHH GSDSEVNQEA KPEVKPEVKP ETHINLKVSD GSSEIFFKIK KTTPLRRLME AFAKRQGKEM DSLRFLYDGI RIQADQTPE DLDMEDNDII EAHREQIGGA WSHPQFEKSA TVSPTRARNM KDFENQITEL KKENFNLKLR IYFLEERMQQ E FHGDDEAA ELMQQVNVLK LTVEDLEKER DFYFGKLRNI ELICQENEGE NDPVLQRIVD ILYATDEGFV I

UniProtKB: Small ubiquitin-related modifier, CDK5 regulatory subunit-associated protein 2, Microtubule-associated protein RP/EB family member 1

Macromolecule #7: Isoform 1 of Tubulin alpha-1B chain

• Macromolecule: Name: Isoform 1 of Tubulin alpha-1B chain / type: protein_or_peptide / ID: 7 / Number of copies: 11 / Enantiomer: LEVO; EC number: Hydrolases; Acting on acid anhydrides; Acting on GTP to facilitate cellular and subcellular movement
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 50.190418 KDa
• Recombinant expression: Organism: Spodoptera frugiperda (fall armyworm)

Sequence

String:

MRECISIHVG QAGVQIGNAC WELYCLEHGI QPDGQMPSDK TIGGGDDSFN TFFSETGAGK HVPRAVFVDL EPTVIDEVRT GTYRQLFHP EQLITGKEDA ANNYARGHYT IGKEIIDLVL DRIRKLADQC TGLQGFLVFH SFGGGTGSGF TSLLMERLSV D YGKKSKLE FSIYPAPQVS TAVVEPYNSI LTTHTTLEHS DCAFMVDNEA IYDICRRNLD IERPTYTNLN RLISQIVSSI TA SLRFDGA LNVDLTDFQT NLVPYPRIHF PLATYAPVIS AEKAYHEQLS VAEITNACFE PANQMVKCDP RHGKYMACCL LYR GDVVPK DVNAAIATIK TKRSIQFVDW CPTGFKVGIN YQPPTVVPGG DLAKVQRAVC MLSNTTAIAE AWARLDHKFD LMYA KRAFV HWYVGEGMEE GEFSEAREDM AALEKDYEEV GVDSVEGEGE EEGEEY

UniProtKB: Tubulin alpha-1B chain

Macromolecule #8: Tubulin beta-3 chain

• Macromolecule: Name: Tubulin beta-3 chain / type: protein_or_peptide / ID: 8 / Number of copies: 11 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 50.906703 KDa
• Recombinant expression: Organism: Spodoptera frugiperda (fall armyworm)

Sequence

String:

MREIVHIQAG QCGNQIGAKF WEVISDEHGI DPSGNYVGDS DLQLERISVY YNEASSHKYV PRAILVDLEP GTMDSVRSGA FGHLFRPDN FIFGQSGAGN NWAKGHYTEG AELVDSVLDV VRKECENCDC LQGFQLTHSL GGGTGSGMGT LLISKVREEY P DRIMNTFS VVPSPKVSDT VVEPYNATLS IHQLVENTDE TYCIDNEALY DICFRTLKLA TPTYGDLNHL VSATMSGVTT SL RFPGQLN ADLRKLAVNM VPFPRLHFFM PGFAPLTARG SQQYRALTVP ELTQQMFDAK NMMAACDPRH GRYLTVATVF RGR MSMKEV DEQMLAIQSK NSSYFVEWIP NNVKVAVCDI PPRGLKMSST FIGNSTAIQE LFKRISEQFT AMFRRKAFLH WYTG EGMDE MEFTEAESNM NDLVSEYQQY QDATAEEEGE MYEDDEEESE AQGPK(GDP)

UniProtKB: Tubulin beta-3 chain

Macromolecule #9: Mitotic-spindle organizing protein 2A

• Macromolecule: Name: Mitotic-spindle organizing protein 2A / type: protein_or_peptide / ID: 9 / Number of copies: 4 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 16.240576 KDa

Sequence

String:

MAAQGVGPGP GSAAPPGLEA ARQKLALRRK KVLSTEEMEL YELAQAAGGG IDPDVFKILV DLLKLNVAPL AVFQMLKSMC AGQRLASEP QDPAAVSLPT SSVPETRGRD KGSAALGGVL ALAERSNHEG SSQRMPRQPS ATRLPKGGGP GKSPTQGST

UniProtKB: Mitotic-spindle organizing protein 2A

Macromolecule #10: Gamma-tubulin complex component 4

• Macromolecule: Name: Gamma-tubulin complex component 4 / type: protein_or_peptide / ID: 10 / Number of copies: 2 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 76.179969 KDa

Sequence

String:

MIHELLLALS GYPGSIFTWN KRSGLQVSQD FPFLHPSETS VLNRLCRLGT DYIRFTEFIE QYTGHVQQQD HHPSQQGQGG LHGIYLRAF CTGLDSVLQP YRQALLDLEQ EFLGDPHLSI SHVNYFLDQF QLLFPSVMVV VEQIKSQKIH GCQILETVYK H SCGGLPPV RSALEKILAV CHGVMYKQLS AWMLHGLLLD QHEEFFIKQG PSSGNVSAQP EEDEEDLGIG GLTGKQLREL QD LRLIEEE NMLAPSLKQF SLRVEILPSY IPVRVAEKIL FVGESVQMFE NQNVNLTRKG SILKNQEDTF AAELHRLKQQ PLF SLVDFE QVVDRIRSTV AEHLWKLMVE ESDLLGQLKI IKDFYLLGRG ELFQAFIDTA QHMLKTPPTA VTEHDVNVAF QQSA HKVLL DDDNLLPLLH LTIEYHGKEH KADATQAREG PSRETSPREA PASGWAALGL SYKVQWPLHI LFTPAVLEKY NVVFK YLLS VRRVQAELQH CWALQMQRKH LKSNQTDAIK WRLRNHMAFL VDNLQYYLQV DVLESQFSQL LHQINSTRDF ESIRLA HDH FLSNLLAQSF ILLKPVFHCL NEILDLCHSF CSLVSQNLGP LDERGAAQLS ILVKGFSRQS SLLFKILSSV RNHQINS DL AQLLLRLDYN KYYTQAGGTL GSFGM

UniProtKB: Gamma-tubulin complex component 4

Macromolecule #11: Gamma-tubulin complex component 5

• Macromolecule: Name: Gamma-tubulin complex component 5 / type: protein_or_peptide / ID: 11 / Number of copies: 2 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 118.467547 KDa

Sequence

String:

MARHGPPWSR LDAQQERDVR ELVRGVAGLQ DEADPNFQLA LNFAWSNFRF HRFLDVNSHK IEKTIEGIYE KFVIHSDLSK AASWKRLTE EFLNAPLPSI KEIKTDAHYS ILSLLLCLSD SPSNSSYVET PRNKEVEKKD DFDWGKYLME DEEMDIGPYM D TPNWSEES EEENDQQPLS REDSGIQVDR TPLEEQDQNR KLDPCISWKD EPDDRSWLEH HVVHQYWTAR PSQFPHSLHL HS NLAAVWD QHLYSSDPLY VPDDRVLVTE TQVIRETLWL LSGVKKLFIF QLIDGKVTVR NNIIVTHLTH SCLRSVLEQI AAY GQVVFR LQEFIDEVMG HSSESMLPGS GSVPKKSTEA PFRTYQAFMW ALYKYFISFK EELAEIEKCI INNDTTITLA IVVD KLAPR LSQLKVLHKV FSTGVAEVPP DTRNVVRASH LLNTLYKAIL EYDNVGEASE QTVSLLFSLW VETVRPYLQT VDEWI VHGH LWDGAREFII QRNKNVPVNH RDFWYATYTL YSVSEKTENE EKMSDNASAS SGSDQGPSSR QHTMVSFLKP VLKQII MAG KSMQLLKNLQ CAESTTCQAG ARDAERKSLY TLFLESVQSR LRHGEDSTPQ VLTEQQATKE NLMKMQSIAE SHLELDD VH DPLLAINFAR MYLEQSDFHE KFAGGDVCVD RSSESVTCQT FELTLRSCLY PHIDKQYLDC CGNLMQTLKK DYRLVEYL Q AMRNFFLMEG GDTMYDFYTS IFDKIREKET WQNVSFLNVQ LQEAVGQRYP EDSSRLSISF ENVDTAKKKL PVHILDGLT LSYKVPWPVD IVISLECQKI YNQVFLLLLQ IKWAKYSLDV LLFGELVSTA EKPRLKEGLI HEQDTVAQFG PQKEPVRQQI HRMFLLRVK LMHFVNSLHN YIMTRILHST GLEFQHQVEE AKDLDQLIKI HYRYLSTIHD RCLLREKVSF VKEAIMKVLN L ALMFADGW QAGLGTWRME SIEKMESDFK NCHMFLVTIL NKAVCRGSFP HLESLALSLM AGMEQS

UniProtKB: Gamma-tubulin complex component 5

Macromolecule #12: Tubulin gamma-1 chain

• Macromolecule: Name: Tubulin gamma-1 chain / type: protein_or_peptide / ID: 12 / Number of copies: 14 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 51.22777 KDa

Sequence

String:

MPREIITLQL GQCGNQIGFE FWKQLCAEHG ISPEGIVEEF ATEGTDRKDV FFYQADDEHY IPRAVLLDLE PRVIHSILNS PYAKLYNPE NIYLSEHGGG AGNNWASGFS QGEKIHEDIF DIIDREADGS DSLEGFVLCH SIAGGTGSGL GSYLLERLND R YPKKLVQT YSVFPNQDEM SDVVVQPYNS LLTLKRLTQN ADCVVVLDNT ALNRIATDRL HIQNPSFSQI NQLVSTIMSA ST TTLRYPG YMNNDLIGLI ASLIPTPRLH FLMTGYTPLT TDQSVASVRK TTVLDVMRRL LQPKNVMVST GRDRQTNHCY IAI LNIIQG EVDPTQVHKS LQRIRERKLA NFIPWGPASI QVALSRKSPY LPSAHRVSGL MMANHTSISS LFERTCRQYD KLRK REAFL EQFRKEDMFK DNFDEMDTSR EIVQQLIDEY HAATRPDYIS WGTQEQ

UniProtKB: Tubulin gamma-1 chain

Macromolecule #13: GUANOSINE-5'-TRIPHOSPHATE

• Macromolecule: Name: GUANOSINE-5'-TRIPHOSPHATE / type: ligand / ID: 13 / Number of copies: 11 / Formula: GTP
• Molecular weight: Theoretical: 523.18 Da

Chemical component information

ChemComp-GTP:
GUANOSINE-5'-TRIPHOSPHATE / GTP, energy-carrying molecule*YM

Experimental details

Structure determination

• Method: cryo EM
• Processing: single particle reconstruction
• Aggregation state: particle

Sample preparation

• Buffer: pH: 6.9
• Grid: Model: Quantifoil R1.2/1.3 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 45 sec. / Pretreatment - Atmosphere: AIR / Details: 15 mA
• Vitrification: Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 310 K / Instrument: FEI VITROBOT MARK IV

Electron microscopy

• Microscope: TFS KRIOS
• Software: Name: EPU
• Image recording: Film or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Number real images: 23663 / Average exposure time: 1.3 sec. / Average electron dose: 40.01 e/Ų
• Electron beam: Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
• Electron optics: C2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 81000
• Sample stage: Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
• Experimental equipment: Model: Titan Krios / Image courtesy: FEI Company

Image processing

• Particle selection: Number selected: 1776484
• CTF correction: Software - Name: CTFFIND (ver. 4) / Type: PHASE FLIPPING AND AMPLITUDE CORRECTION
• Startup model: Type of model: INSILICO MODEL / In silico model: CryoSPARC Ab Initio Reconstruction
• Final reconstruction: Applied symmetry - Point group: C₁ (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.67 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 5.0) / Number images used: 652699
• Initial angle assignment: Type: ANGULAR RECONSTITUTION / Software: (Name: cryoSPARC (ver. 4.7.0), RELION (ver. 5.0))
• Final angle assignment: Type: ANGULAR RECONSTITUTION / Software - Name: RELION (ver. 5.0)
• Final 3D classification: Details: Several 3D classification and heterogeneity analysis methods

Atomic model buiding 1

Initial model

PDB ID	Chain
7as4	source_name: PDB, initial_model_type: experimental model
6x0u	source_name: PDB, initial_model_type: experimental model

• Refinement: Space: REAL / Protocol: FLEXIBLE FIT

Output model

PDB-9smx:
CM1-activated gTuRC in complex with nascent alpha-E254D mutant microtubules