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- EMDB-70135: The KICSTOR-GATOR1 complex -

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Basic information

Entry
Database: EMDB / ID: EMD-70135
TitleThe KICSTOR-GATOR1 complex
Map dataComposite map of the KICSTOR:GATOR1 complex
Sample
  • Complex: Binary complex of KICSTOR-GATOR1
    • Complex: GATOR1 subcomplex
      • Protein or peptide: GATOR complex protein NPRL2
      • Protein or peptide: GATOR1 complex protein DEPDC5
      • Protein or peptide: GATOR complex protein NPRL3
    • Complex: KICSTOR subcomplex
      • Protein or peptide: KICSTOR complex protein SZT2
      • Protein or peptide: KICSTOR complex protein kaptin
      • Protein or peptide: KICSTOR subunit 2
      • Protein or peptide: KICSTOR complex protein ITFG2
KeywordsLysosome / GATOR1 / KICSTOR / cell growth / amino acid sensing / mTOR / KPTN / ITFG2 / C12orf66 / SZT2 / NPRL2 / NPRL3 / DEPDC5 / CELL CYCLE
Function / homology
Function and homology information


regulation of superoxide dismutase activity / KICSTOR complex / GATOR1 complex / negative regulation of kinase activity / corpus callosum morphogenesis / aorta morphogenesis / germinal center B cell differentiation / protein localization to lysosome / regulation of TOR signaling / Amino acids regulate mTORC1 ...regulation of superoxide dismutase activity / KICSTOR complex / GATOR1 complex / negative regulation of kinase activity / corpus callosum morphogenesis / aorta morphogenesis / germinal center B cell differentiation / protein localization to lysosome / regulation of TOR signaling / Amino acids regulate mTORC1 / stereocilium / postsynaptic actin cytoskeleton / cardiac muscle tissue development / vacuolar membrane / ventricular septum development / pigmentation / roof of mouth development / cellular response to glucose starvation / intercellular bridge / negative regulation of TORC1 signaling / positive regulation of autophagy / GTPase activator activity / cellular response to amino acid starvation / actin filament organization / central nervous system development / post-embryonic development / small GTPase binding / actin filament binding / peroxisome / lamellipodium / lysosome / intracellular signal transduction / intracellular membrane-bounded organelle / lysosomal membrane / protein-containing complex binding / perinuclear region of cytoplasm / glutamatergic synapse / Golgi apparatus / nucleoplasm / cytosol
Similarity search - Function
KICSTOR subunit 2 / KICSTOR complex protein C12orf66-like, central domain superfamily / KICSTOR complex C12orf66 like / Kaptin / Integrin-alpha FG-GAP repeat-containing protein 2 / Integrin-alpha FG-GAP repeat-containing protein 2 / Protein SZT2 / : / IML1 N-terminal double psi beta barrel domain / Nitrogen permease regulator 3 ...KICSTOR subunit 2 / KICSTOR complex protein C12orf66-like, central domain superfamily / KICSTOR complex C12orf66 like / Kaptin / Integrin-alpha FG-GAP repeat-containing protein 2 / Integrin-alpha FG-GAP repeat-containing protein 2 / Protein SZT2 / : / IML1 N-terminal double psi beta barrel domain / Nitrogen permease regulator 3 / Nitrogen permease regulator 2 / : / Nitrogen Permease regulator of amino acid transport activity 3 / Nitrogen permease regulator 2 / GATOR1 complex protein NPRL3, C-terminal HTH / Vacuolar membrane-associated protein Iml1 / DEPDC5 protein, C-terminal / : / Vacuolar membrane-associated protein Iml1, N-terminal domain / DEPDC5 protein C-terminal region / Domain found in Dishevelled, Egl-10, and Pleckstrin (DEP) / DEP domain profile. / Domain found in Dishevelled, Egl-10, and Pleckstrin / DEP domain / Integrin alpha, N-terminal / Winged helix DNA-binding domain superfamily / Winged helix-like DNA-binding domain superfamily / WD40-repeat-containing domain superfamily
Similarity search - Domain/homology
GATOR1 complex protein DEPDC5 / GATOR1 complex protein NPRL3 / KICSTOR complex protein SZT2 / GATOR1 complex protein NPRL2 / KICSTOR complex protein ITFG2 / KICSTOR subunit 2 / KICSTOR complex protein kaptin
Similarity search - Component
Biological speciesHomo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.2 Å
AuthorsBayly-Jones C / Lupton CJ / Chang YG / Ellisdon AM
Funding support Australia, 4 items
OrganizationGrant numberCountry
National Health and Medical Research Council (NHMRC, Australia)APP2036864 Australia
Australian Research Council (ARC)DE240100992 Australia
Other governmentMCRF21036 Australia
Australian Research Council (ARC)LE170100016 Australia
CitationJournal: Cell / Year: 2026
Title: Structure of the lysosomal KICSTOR-GATOR1-SAMTOR nutrient-sensing supercomplex.
Authors: Christopher J Lupton / Charles Bayly-Jones / Shuqi Dong / Terrance Lam / Wentong Luo / Gareth D Jones / Chantel Mastos / Nicholas J Frescher / San S Lim / Alastair C Keen / Luke E Formosa / ...Authors: Christopher J Lupton / Charles Bayly-Jones / Shuqi Dong / Terrance Lam / Wentong Luo / Gareth D Jones / Chantel Mastos / Nicholas J Frescher / San S Lim / Alastair C Keen / Luke E Formosa / Hari Venugopal / Yong-Gang Chang / Michelle L Halls / Andrew M Ellisdon /
Abstract: The guanosine triphosphate (GTP)-bound state of the heterodimeric Rag GTPases functions as a molecular switch regulating mechanistic target of rapamycin complex 1 (mTORC1) activation at the lysosome ...The guanosine triphosphate (GTP)-bound state of the heterodimeric Rag GTPases functions as a molecular switch regulating mechanistic target of rapamycin complex 1 (mTORC1) activation at the lysosome downstream of amino acid fluctuations. Under low amino acid conditions, GTPase-activating protein (GAP) activity toward Rags 1 (GATOR1) promotes RagA GTP hydrolysis, preventing mTORC1 activation. KICSTOR recruits and regulates GATOR1 at the lysosome by undefined mechanisms. Here, we resolve the KICSTOR-GATOR1 structure, revealing a striking ∼60-nm crescent-shaped assembly. GATOR1 anchors to KICSTOR via an extensive interface, and mutations that disrupt this interaction impair mTORC1 regulation. The S-adenosylmethionine sensor SAMTOR binds KICSTOR in a manner incompatible with metabolite binding, providing structural insight into methionine sensing via SAMTOR-KICSTOR association. We discover that KICSTOR and GATOR1 form a dimeric supercomplex. This assembly restricts GATOR1 to an orientation that favors the low-affinity active GAP mode of Rag GTPase engagement while sterically restricting access to the high-affinity inhibitory mode, consistent with a model of an active lysosomal GATOR1 docking complex.
History
DepositionApr 9, 2025-
Header (metadata) releaseFeb 18, 2026-
Map releaseFeb 18, 2026-
UpdateFeb 18, 2026-
Current statusFeb 18, 2026Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_70135.png

Downloads & links

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Map

FileDownload / File: emd_70135.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationComposite map of the KICSTOR:GATOR1 complex
Projections & slices

Image control

Size
Brightness
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Others
AxesZ (Sec.)Y (Row.)X (Col.)
1 Å/pix.
x 480 pix.
= 480. Å		1 Å/pix.
x 480 pix.
= 480. Å		1 Å/pix.
x 480 pix.
= 480. Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 1.8
Minimum - Maximum0.0 - 20.187868000000002
Average (Standard dev.)0.026306354 (±0.38450256)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions480480480
Spacing480480480
CellA=B=C: 480.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Binary complex of KICSTOR-GATOR1

EntireName: Binary complex of KICSTOR-GATOR1
Components
  • Complex: Binary complex of KICSTOR-GATOR1
    • Complex: GATOR1 subcomplex
      • Protein or peptide: GATOR complex protein NPRL2
      • Protein or peptide: GATOR1 complex protein DEPDC5
      • Protein or peptide: GATOR complex protein NPRL3
    • Complex: KICSTOR subcomplex
      • Protein or peptide: KICSTOR complex protein SZT2
      • Protein or peptide: KICSTOR complex protein kaptin
      • Protein or peptide: KICSTOR subunit 2
      • Protein or peptide: KICSTOR complex protein ITFG2

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Supramolecule #1: Binary complex of KICSTOR-GATOR1

SupramoleculeName: Binary complex of KICSTOR-GATOR1 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 298 KDa

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Supramolecule #2: GATOR1 subcomplex

SupramoleculeName: GATOR1 subcomplex / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1-#2, #7
Source (natural)Organism: Homo sapiens (human)

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Supramolecule #3: KICSTOR subcomplex

SupramoleculeName: KICSTOR subcomplex / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #3-#6
Source (natural)Organism: Homo sapiens (human)

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Macromolecule #1: GATOR complex protein NPRL2

MacromoleculeName: GATOR complex protein NPRL2 / type: protein_or_peptide / ID: 1 / Details: N-terminal HA tagged NPRL2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 45.799547 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MGYPYDVPDY ADLNGGGGGS TMGSGCRIEC IFFSEFHPTL GPKITYQVPE DFISRELFDT VQVYIITKPE LQNKLITVTA MEKKLIGCP VCIEHKKYSR NALLFNLGFV CDAQAKTCAL EPIVKKLAGY LTTLELESSF VSMEESKQKL VPIMTILLEE L NASGRCTL ...String:
MGYPYDVPDY ADLNGGGGGS TMGSGCRIEC IFFSEFHPTL GPKITYQVPE DFISRELFDT VQVYIITKPE LQNKLITVTA MEKKLIGCP VCIEHKKYSR NALLFNLGFV CDAQAKTCAL EPIVKKLAGY LTTLELESSF VSMEESKQKL VPIMTILLEE L NASGRCTL PIDESNTIHL KVIEQRPDPP VAQEYDVPVF TKDKEDFFNS QWDLTTQQIL PYIDGFRHIQ KISAEADVEL NL VRIAIQN LLYYGVVTLV SILQYSNVYC PTPKVQDLVD DKSLQEACLS YVTKQGHKRA SLRDVFQLYC SLSPGTTVRD LIG RHPQQL QHVDERKLIQ FGLMKNLIRR LQKYPVRVTR EEQSHPARLY TGCHSYDEIC CKTGMSYHEL DERLENDPNI IICW K

UniProtKB: GATOR1 complex protein NPRL2

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Macromolecule #2: GATOR1 complex protein DEPDC5

MacromoleculeName: GATOR1 complex protein DEPDC5 / type: protein_or_peptide / ID: 2 / Details: N-terminal FLAG tagged DEPDC5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 183.207703 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MGDYKDDDDK GGGGGASTMR TTKVYKLVIH KKGFGGSDDE LVVNPKVFPH IKLGDIVEIA HPNDEYSPLL LQVKSLKEDL QKETISVDQ TVTQVFRLRP YQDVYVNVVD PKDVTLDLVE LTFKDQYIGR GDMWRLKKSL VSTCAYITQK VEFAGIRAQA G ELWVKNEK ...String:
MGDYKDDDDK GGGGGASTMR TTKVYKLVIH KKGFGGSDDE LVVNPKVFPH IKLGDIVEIA HPNDEYSPLL LQVKSLKEDL QKETISVDQ TVTQVFRLRP YQDVYVNVVD PKDVTLDLVE LTFKDQYIGR GDMWRLKKSL VSTCAYITQK VEFAGIRAQA G ELWVKNEK VMCGYISEDT RVVFRSTSAM VYIFIQMSCE MWDFDIYGDL YFEKAVNGFL ADLFTKWKEK NCSHEVTVVL FS RTFYDAK SVDEFPEINR ASIRQDHKGR FYEDFYKVVV QNERREEWTS LLVTIKKLFI QYPVLVRLEQ AEGFPQGDNS TSA QGNYLE AINLSFNVFD KHYINRNFDR TGQMSVVITP GVGVFEVDRL LMILTKQRMI DNGIGVDLVC MGEQPLHAVP LFKL HNRSA PRDSRLGDDY NIPHWINHSF YTSKSQLFCN SFTPRIKLAG KKPASEKAKN GRDTSLGSPK ESENALPIQV DYDAY DAQV FRLPGPSRAQ CLTTCRSVRE RESHSRKSAS SCDVSSSPSL PSRTLPTEEV RSQASDDSSL GKSANILMIP HPHLHQ YEV SSSLGYTSTR DVLENMMEPP QRDSSAPGRF HVGSAESMLH VRPGGYTPQR ALINPFAPSR MPMKLTSNRR RWMHTFP VG PSGEAIQIHH QTRQNMAELQ GSGQRDPTHS SAELLELAYH EAAGRHSNSR QPGDGMSFLN FSGTEELSVG LLSNSGAG M NPRTQNKDSL EDSVSTSPDP ILTLSAPPVV PGFCCTVGVD WKSLTTPACL PLTTDYFPDR QGLQNDYTEG CYDLLPEAD IDRRDEDGVQ MTAQQVFEEF ICQRLMQGYQ IIVQPKTQKP NPAVPPPLSS SPLYSRGLVS RNRPEEEDQY WLSMGRTFHK VTLKDKMIT VTRYLPKYPY ESAQIHYTYS LCPSHSDSEF VSCWVEFSHE RLEEYKWNYL DQYICSAGSE DFSLIESLKF W RTRFLLLP ACVTATKRIT EGEAHCDIYG DRPRADEDEW QLLDGFVRFV EGLNRIRRRH RSDRMMRKGT AMKGLQMTGP IS THSLEST APPVGKKGTS ALSALLEMEA SQKCLGEQQA AVHGGKSSAQ SAESSSVAMT PTYMDSPRKD GAFFMEFVRS PRT ASSAFY PQVSVDQTAT PMLDGTSLGI CTGQSMDRGN SQTFGNSQNI GEQGYSSTNS SDSSSQQLVA SSLTSSSTLT EILE AMKHP STGVQLLSEQ KGLSPYCFIS AEVVHWLVNH VEGIQTQAMA IDIMQKMLEE QLITHASGEA WRTFIYGFYF YKIVT DKEP DRVAMQQPAT TWHTAGVDDF ASFQRKWFEV AFVAEELVHS EIPAFLLPWL PSRPASYASR HSSFSRSFGG RSQAAA LLA ATVPEQRTVT LDVDVNNRTD RLEWCSCYYH GNFSLNAAFE IKLHWMAVTA AVLFEMVQGW HRKATSCGFL LVPVLEG PF ALPSYLYGDP LRAQLFIPLN ISCLLKEGSE HLFDSFEPET YWDRMHLFQE AIAHRFGFVQ DKYSASAFNF PAENKPQY I HVTGTVFLQL PYSKRKFSGQ QRRRRNSTSS TNQNMFCEER VGYNWAYNTM LTKTWRSSAT GDEKFADRLL KDFTDFCIN RDNRLVTFWT SCLEKMHASA P

UniProtKB: GATOR1 complex protein DEPDC5

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Macromolecule #3: KICSTOR complex protein SZT2

MacromoleculeName: KICSTOR complex protein SZT2 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 380.3865 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MEMDYKDDDD KGGGGGASTM ASERPEPEVE EAGQVFLLMK KDYRISRNVR LAWFLSHLHQ TVQATPQEML LQSEQELEVL SVLPPGWQP DEPVVPRPFL LVPSTRVTFL AWQYRFVIEL DLSPSTGIVD DSTGEILFDE VFHALSRCLG GLLRPFRVPG S CIDFQPEI ...String:
MEMDYKDDDD KGGGGGASTM ASERPEPEVE EAGQVFLLMK KDYRISRNVR LAWFLSHLHQ TVQATPQEML LQSEQELEVL SVLPPGWQP DEPVVPRPFL LVPSTRVTFL AWQYRFVIEL DLSPSTGIVD DSTGEILFDE VFHALSRCLG GLLRPFRVPG S CIDFQPEI YVTIQAYSSI IGLQSHQVLV QGCLLDPSQR EVFLQQIYEQ LCLFEDKVAT MLQQQYDPQS QAEDQSPDSG DL LGRKVGV SMVTADLGLV SMIRQGILAL QLLPSNSSAG IIVITDGVTS VPDVAVCETL LNQLRSGTVA CSFVQVGGVY SYD CSFGHV PNVELMKFIA MATFGSYLST CPEPEPGNLG LTVYHRAFLL YSFLRSGEAL NPEYYCGSQH RLFNEHLVSA SSNP ALALR RKKHTEKEVP ADLVSTVSVR LREGYSVREV TLAKGGSQLE VKLVLLWKHN MRIEYVAMAP WPLEPEGPRV TRVEV TMEG GYDILHDVSC ALRQPIRSLY RTHVIRRFWN TLQSINQTDQ MLAHLQSFSS VPEHFTLPDS TKSGVPLFYI PPGSTT PVL SLQPSGSDSS HAQFAAYWKP VLSMDANSWQ RWLHMHRLVL ILEHDTPIPK HLHTPGSNGR YSTIQCRISH SSLTSLL RD WSSFVLVEGY SYVKLLSSAP DQPPNSFYMV RIISKAPCMV LRLGFPIGTP APARHKIVSG LREEILRLRF PHRVQSKE P TPKVKRKGLG GAGGGSSPSK SPPVLGPQQA LSDRPCLVVL HKPLDKLLIR YEKLPLDYRA PFLLTLEPPG PLPLVSGRS ASSSLASLSR YLYHQRWLWS VPSGLAPALP LSAIAQLLSI LTEVRLSEGF HFACSGEGII NMVLELPIQN EPPGQAAAEE KHTCVVQYI LFPPHSTSTK DSFSTDDDND VEVEALEGDS ELNLVTEVWV EPQYGRVGPG PGIWKHLQDL TYSEIPQALH P RDAACIGS MLSFEYLIQL CQSKEWGPLP PEPRVSDGLD QGGDTCVHEI PFHFDLMGLL PQCQQLQMFF LLLAREPEGV PF AEGSCPA NDMVLCLLHS CLGQELSDRE IPLTPVDQAA FLSEVLRRTC HVPGAEGPLL GVHGIPKEQA VGSTQATGDS AFT SLSVGL PETLKPLISA QPPQWRCYAR LVNPQHVFLT FLPATFSDVQ RLAACGLEGP PQEETKPKFG DWSGAPSLKD LGGT GIKAT KSHVPVLSVT LASDNAQNQG ELSPPFRRDL QAYAGRQASQ TESADGPRTR CPVYIYSCSL EALREQMVGM QPPQA PRDL IFRTQFLDHP SPSSAWMEPR YKEAANHCAL LQEHAQRCYV RGLFRSLQQA QSVTSQDLLT AVDACEELLQ EIDITP FLL ALCGHTWGLP HAPPSPGPLS PGPFSSSMEE GAEPRERAIL ASESSIETED LSEPEFQSTR VPGIPDPGPE ISLTDVC QL RGEAHGALHS VIQEKFLEIS RLHFRTVPSN PHYFFYCPPS SRREDEGPRD TVDRKISDLE FSEAELMGEE GDTSACCV V TESDPELEVE YRESRESDLG PAGLDSASLS DVDTVNPDED SFSILGGDSP TGPESFLHDL PPLFLHLTCS VRLRGQHSS VPVCSLPTCL GQVLSSLEGP PVGGRVPLRD LSVTLDVFML TLPLEVELPT ASDPQHHRST SESSASFPRS PGQPSSLRSD DGLGPPLPP PEEERHPGLS NLATPHRLAI ETTMNEIRWL LEDEMVGALR RGGIPQSPAL HRAAAHIHSS PGRSTCLRQT L PLSFVFGP ERSLTQFKEE FRRLHLPGHV LLEDPDSGFF FVAAGQQPGG SHGEPSSAAW AWHSHEDRAE GIEGETLTAS PQ APGSPED SEGVPLISLP RVPQGGSQPG PSRGLSLMSS QGSVDSDHLG YDGGSSGSDS EGPNDTLGEK APFTLRTPPG PAP PQPSLS GLPGPCLPDF WLIVRVLQDR VEVYAHARSL IREDGGPGTE CRHLQQLLVR RVGEICREVN QRLLLQDLHD SHVC NSLLV AESEEDLWRS ETPFHSRQRA PLPSDDYAAD ESCAPRGYLA ATMQFVPGHF SCDVVWGTVI RVHSRLKMGP SMGVS RAIQ ALRSVLNAFS VVNRKNMFVY QERATKAVYY LRLLETSCSD RPWKGDALPP SLALSRSQEP IYSEEASGPR SPLDMV SSR SSDAARPVGQ VDRHIQLLVH GVGQAGPEIT DELVRVLCRR LDEATLDVIT VMLVRNCKLT PADVEFIQPP GSLPSEV LH LALPTSCRPW LPALAWYLRQ NLLIFLHSPK YTDSNSRNHF QHPLPPQGGL PDLDIYLYNK PGGQGTGGKG VACITLAF V DEGGAPLSLA LWPPSSPGPP DPLREEEFEQ LTQVIRCPVV VDSSSAQNGA PRLRLDVWEK GNISIVQLEE KLRGAARQA LADAIIELQL LPASLCTEDT PTGSLRNGSL ETKSSAGRAS TFPPAPVPGE PVTPPSKAGR RSFWDMLSKT ECGDLGSPKT TDDIVLDRP EDTRGRRRHK TESVRTPGGA ERAPGSDSGA QRQKRRTTQL EEGEVGTLHP VFARVAQRWM EFMVQIGCAS V SRSSAHMV SRFLLPSILS EFTALVTSMA GDTSVRIFEQ HLGSEPEIFG PCSPGQLGPS PRPAAERHLL LLGRNFLQWR RP TQQAAKA MQRFEPGGDG SSGRNAPRQR LLLLEVVDKK LQLLTYNWAP DLGAALGRAL VRLVQWQNAR AHLIFCLLSQ KLG LFHHYG QLDFPVRDEK EPNPFLLPTM EVETLIRSAS PPLSREQGRL SGSSRGGGPL PLDTFPFDEA LRDITAARPS SVLG PVPRP PDPVTYHGQQ FLEIKMAERR ELERQMKMEN LFVTWQQRST PATMPISAGE LETLKQSSRL VHYCATAMLF DPAAW LHGP PETSGPPDGQ RRHRPESGSG SREAPTSCES LDVSPPGARE EPWLKELSLA FLQQYVQYLQ SIGFVLVPLR PPSPAR STS RPRAMAILGT EGRGSFSCPK TKTDGSPKST SSPVTTYHLQ RALPGGIILM ELAFQGCYFC VKQFALECSR IPMGQAV NS QLSMLFTEEC DKVRDLMHVH SFSYDFHLRL VHQHVLGAHL VLRHGYHLTT FLRHFLAHHP DGPHFGRNHI YQGTLELP T PLIAAHQLYN YVADHASSYH MKPLRMARPG GPEHNEYALV SAWHSSGSYL DSEGLRHQDD FDVSLLVCHC AAPFEEQGE AERHVLRLQF FVVLTSQREL FPRLTADMRR FRKPPRLPPE PEAPGSSAGS PGEASGLILA PGPAPLFPPL AAEVGMARAR LAQLVRLAG GHCRRDTLWK RLFLLEPPGP DRLRLGGRLA LAELEELLEA VHAKSIGDID PQLDCFLSMT VSWYQSLIKV L LSRFPQSC RHFQSPDLGT QYLVVLNQKF TDCFVLVFLD SHLGKTSLTV VFREPFPVQP QDSESPPAQL VSTYHHLESV IN TACFTLW TRLL

UniProtKB: KICSTOR complex protein SZT2

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Macromolecule #4: KICSTOR complex protein kaptin

MacromoleculeName: KICSTOR complex protein kaptin / type: protein_or_peptide / ID: 4 / Details: N-terminal HA tagged KPTN / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 50.216473 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MGYPYDVPDY ADLNGGGGGS TMMGEAAVAA GPCPLREDSF TRFSSQSNVY GLAGGAGGRG ELLAATLKGK VLGFRYQDLR QKIRPVAKE LQFNYIPVDA EIVSIDTFNK SPPKRGLVVG ITFIKDSGDK GSPFLNIYCD YEPGSEYNLD SIAQSCLNLE L QFTPFQLC ...String:
MGYPYDVPDY ADLNGGGGGS TMMGEAAVAA GPCPLREDSF TRFSSQSNVY GLAGGAGGRG ELLAATLKGK VLGFRYQDLR QKIRPVAKE LQFNYIPVDA EIVSIDTFNK SPPKRGLVVG ITFIKDSGDK GSPFLNIYCD YEPGSEYNLD SIAQSCLNLE L QFTPFQLC HAEVQVGDQL ETVFLLSGND PAIHLYKENE GLHQFEEQPV ENLFPELTNL TSSVLWLDVH NFPGTSRRLS AL GCQSGYV RVAHVDQRSR EVLQMWSVLQ DGPISRVIVF SLSAAKETKD RPLQDEYSVL VASMLEPAVV YRDLLNRGLE DQL LLPGSD QFDSVLCSLV TDVDLDGRPE VLVATYGQEL LCYKYRGPES GLPEAQHGFH LLWQRSFSSP LLAMAHVDLT GDGL QELAV VSLKGVHILQ HSLIQASELV LTRLRHQVEQ RRRRLQGLED GAGAGPAENA AS

UniProtKB: KICSTOR complex protein kaptin

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Macromolecule #5: KICSTOR subunit 2

MacromoleculeName: KICSTOR subunit 2 / type: protein_or_peptide / ID: 5 / Details: N-terminal HA tagged C12orf66 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 52.57393 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MGYPYDVPDY ADLNGGGGGS TMGESIPLAA PVPVEQAVLE TFFSHLGIFS YDKAKDNVEK EREANKSAGG SWLSLLAALA HLAAAEKVY HSLTYLGQKL GGQSFFSRKD SIRTIYTSLH NELKKVVTGR GALGGTAPHV EELLSHLSEQ LCFFVQARME I ADFYEKMY ...String:
MGYPYDVPDY ADLNGGGGGS TMGESIPLAA PVPVEQAVLE TFFSHLGIFS YDKAKDNVEK EREANKSAGG SWLSLLAALA HLAAAEKVY HSLTYLGQKL GGQSFFSRKD SIRTIYTSLH NELKKVVTGR GALGGTAPHV EELLSHLSEQ LCFFVQARME I ADFYEKMY TLSTQKFINA EELVGLLDAI MKKYSSRFHH PILSPLESSF QLEVDVLCHL LKAQAQVSEW KFLPSLVNLH SA HTKLQTW GQIFEKQRET KKHLFGGQSQ KAVQPPHLFL WLMKLKNMLL AKFSFYFHEA LSRQTTASEM KTLTAKANPD FFG KISSFI RKYDAANVSL IFDNRGSESF QGHGYHHPHS YREAPKGVDQ YPAVVSLPSD RPVMHWPNVI MIMTDRTSDL NSLE KVVHF YDDKVQSTYF LTRPEPHFTI VIIFESKKSE RDSHFISFLN EVSLALKNPK VFASLKPGSK G

UniProtKB: KICSTOR subunit 2

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Macromolecule #6: KICSTOR complex protein ITFG2

MacromoleculeName: KICSTOR complex protein ITFG2 / type: protein_or_peptide / ID: 6 / Details: N-terminal myc tagged ITFG2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 51.270758 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MEQKLISEED LNGGGGGSTM RSVSYVQRVA LEFSGSLFPH AICLGDVDND TLNELVVGDT SGKVSVYKND DSRPWLTCSC QGMLTCVGV GDVCNKGKNL LVAVSAEGWF HLFDLTPAKV LDASGHHETL IGEEQRPVFK QHIPANTKVM LISDIDGDGC R ELVVGYTD ...String:
MEQKLISEED LNGGGGGSTM RSVSYVQRVA LEFSGSLFPH AICLGDVDND TLNELVVGDT SGKVSVYKND DSRPWLTCSC QGMLTCVGV GDVCNKGKNL LVAVSAEGWF HLFDLTPAKV LDASGHHETL IGEEQRPVFK QHIPANTKVM LISDIDGDGC R ELVVGYTD RVVRAFRWEE LGEGPEHLTG QLVSLKKWML EGQVDSLSVT LGPLGLPELM VSQPGCAYAI LLCTWKKDTG SP PASEGPT DGSRETPAAR DVVLHQTSGR IHNKNVSTHL IGNIKQGHGT ESSGSGLFAL CTLDGTLKLM EEMEEADKLL WSV QVDHQL FALEKLDVTG NGHEEVVACA WDGQTYIIDH NRTVVRFQVD ENIRAFCAGL YACKEGRNSP CLVYVTFNQK IYVY WEVQL ERMESTNLVK LLETKPEYHS LLQELGVDPD DLPVTRALLH QTLYHPDQPP QCAPSSLQDP T

UniProtKB: KICSTOR complex protein ITFG2

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Macromolecule #7: GATOR complex protein NPRL3

MacromoleculeName: GATOR complex protein NPRL3 / type: protein_or_peptide / ID: 7 / Details: N-terminal HA tagged NPRL3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 65.769 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MGYPYDVPDY ADLNGGGGGS TMRDNTSPIS VILVSSGSRG NKLLFRYPFQ RSQEHPASQT SKPRSRYAAS NTGDHADEQD GDSRFSDVI LATILATKSE MCGQKFELKI DNVRFVGHPT LLQHALGQIS KTDPSPKREA PTMILFNVVF ALRANADPSV I NCLHNLSR ...String:
MGYPYDVPDY ADLNGGGGGS TMRDNTSPIS VILVSSGSRG NKLLFRYPFQ RSQEHPASQT SKPRSRYAAS NTGDHADEQD GDSRFSDVI LATILATKSE MCGQKFELKI DNVRFVGHPT LLQHALGQIS KTDPSPKREA PTMILFNVVF ALRANADPSV I NCLHNLSR RIATVLQHEE RRCQYLTREA KLILALQDEV SAMADGNEGP QSPFHHILPK CKLARDLKEA YDSLCTSGVV RL HINSWLE VSFCLPHKIH YAASSLIPPE AIERSLKAIR PYHALLLLSD EKSLLGELPI DCSPALVRVI KTTSAVKNLQ QLA QDADLA LLQVFQLAAH LVYWGKAIII YPLCENNVYM LSPNASVCLY SPLAEQFSHQ FPSHDLPSVL AKFSLPVSLS EFRN PLAPA VQETQLIQMV VWMLQRRLLI QLHTYVCLMA SPSEEEPRPR EDDVPFTARV GGRSLSTPNA LSFGSPTSSD DMTLT SPSM DNSSAELLPS GDSPLNQRMT ENLLASLSEH ERAAILSVPA AQNPEDLRMF ARLLHYFRGR HHLEEIMYNE NTRRSQ LLM LFDKFRSVLV VTTHEDPVIA VFQALLP

UniProtKB: GATOR1 complex protein NPRL3

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

Concentration0.68 mg/mL
BufferpH: 7.4
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 10 sec. / Pretreatment - Atmosphere: AIR
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 278 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeTFS KRIOS
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 1.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.8 µm / Nominal defocus min: 0.5 µm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
Startup modelType of model: OTHER / Details: Ab initio
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.2 Å / Resolution method: OTHER / Software: (Name: RELION, cryoSPARC)
Details: Resolution estimated as the average nominal resolution of constituent focused refinements. See focused reconstructions.
Number images used: 43700
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD

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Atomic model buiding 1

RefinementProtocol: FLEXIBLE FIT
Output model

PDB-9o5a:
The KICSTOR-GATOR1 complex

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