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- EMDB-33528: Cryo-EM structure of TOC-TIC supercomplex from Chlamydomonas rein... -

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Basic information

Entry
Database: EMDB / ID: EMD-33528
TitleCryo-EM structure of TOC-TIC supercomplex from Chlamydomonas reinhardtii
Map data
Sample
  • Complex: a membrane protein complex
    • Protein or peptide: x 14 types
  • Ligand: x 6 types
KeywordsComplex / membrane protein / TRANSLOCASE
Function / homology
Function and homology information


nucleoid organization / chloroplast fission / chloroplast inner membrane / chloroplast outer membrane / chloroplast membrane / chloroplast / protein transport / Hydrolases; Acting on acid anhydrides; Acting on GTP to facilitate cellular and subcellular movement / hydrolase activity / GTP binding ...nucleoid organization / chloroplast fission / chloroplast inner membrane / chloroplast outer membrane / chloroplast membrane / chloroplast / protein transport / Hydrolases; Acting on acid anhydrides; Acting on GTP to facilitate cellular and subcellular movement / hydrolase activity / GTP binding / membrane / metal ion binding
Similarity search - Function
CCB3/YggT / Chloroplast protein import component Tic20 / Protein TIC56 / YGGT family / Chloroplast import apparatus Tic20-like / GYF domain 2 / GYF domain 2 / AIG1-type guanine nucleotide-binding (G) domain / GTPase GIMA/IAN/Toc / AIG1 family ...CCB3/YggT / Chloroplast protein import component Tic20 / Protein TIC56 / YGGT family / Chloroplast import apparatus Tic20-like / GYF domain 2 / GYF domain 2 / AIG1-type guanine nucleotide-binding (G) domain / GTPase GIMA/IAN/Toc / AIG1 family / AIG1-type G domain profile. / Possible plasma membrane-binding motif in junctophilins, PIP-5-kinases and protein kinases. / MORN motif / MORN repeat / Surface antigen D15-like / Bacterial surface antigen (D15) / Omp85 superfamily domain / P-loop containing nucleoside triphosphate hydrolase
Similarity search - Domain/homology
AIG1-type G domain-containing protein / Flagellar associated protein / Uncharacterized protein / Fanciful K+ uptake-b family transporter / Translocase of chloroplast 34 homolog, chloroplastic / Bacterial surface antigen (D15) domain-containing protein / Protein TIC 20 / Uncharacterized protein / Uncharacterized protein / Uncharacterized protein ...AIG1-type G domain-containing protein / Flagellar associated protein / Uncharacterized protein / Fanciful K+ uptake-b family transporter / Translocase of chloroplast 34 homolog, chloroplastic / Bacterial surface antigen (D15) domain-containing protein / Protein TIC 20 / Uncharacterized protein / Uncharacterized protein / Uncharacterized protein / GYF domain-containing protein / Uncharacterized membrane protein ycf78
Similarity search - Component
Biological speciesChlamydomonas reinhardtii (plant)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.77 Å
AuthorsLiu H / Li AJ / Liu ZF
Funding support China, 4 items
OrganizationGrant numberCountry
Chinese Academy of SciencesYSBR-015 China
National Natural Science Foundation of China (NSFC)31925024 China
Chinese Academy of SciencesXDB37020101 China
Ministry of Science and Technology (MoST, China)2017YFA0503702 China
CitationJournal: Nature / Year: 2023
Title: Architecture of chloroplast TOC-TIC translocon supercomplex.
Authors: Hao Liu / Anjie Li / Jean-David Rochaix / Zhenfeng Liu /
Abstract: Chloroplasts rely on the translocon complexes in the outer and inner envelope membranes (the TOC and TIC complexes, respectively) to import thousands of different nuclear-encoded proteins from the ...Chloroplasts rely on the translocon complexes in the outer and inner envelope membranes (the TOC and TIC complexes, respectively) to import thousands of different nuclear-encoded proteins from the cytosol. Although previous studies indicated that the TOC and TIC complexes may assemble into larger supercomplexes, the overall architectures of the TOC-TIC supercomplexes and the mechanism of preprotein translocation are unclear. Here we report the cryo-electron microscopy structure of the TOC-TIC supercomplex from Chlamydomonas reinhardtii. The major subunits of the TOC complex (Toc75, Toc90 and Toc34) and TIC complex (Tic214, Tic20, Tic100 and Tic56), three chloroplast translocon-associated proteins (Ctap3, Ctap4 and Ctap5) and three newly identified small inner-membrane proteins (Simp1-3) have been located in the supercomplex. As the largest protein, Tic214 traverses the inner membrane, the intermembrane space and the outer membrane, connecting the TOC complex with the TIC proteins. An inositol hexaphosphate molecule is located at the Tic214-Toc90 interface and stabilizes their assembly. Four lipid molecules are located within or above an inner-membrane funnel formed by Tic214, Tic20, Simp1 and Ctap5. Multiple potential pathways found in the TOC-TIC supercomplex may support translocation of different substrate preproteins into chloroplasts.
History
DepositionJun 2, 2022-
Header (metadata) releaseJan 11, 2023-
Map releaseJan 11, 2023-
UpdateOct 30, 2024-
Current statusOct 30, 2024Processing site: PDBj / Status: Released

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Structure visualization

Supplemental images

emd_33528.png

Downloads & links

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Map

FileDownload / File: emd_33528.map.gz / Format: CCP4 / Size: 1000 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
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Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.68 Å/pix.
x 640 pix.
= 432. Å		0.68 Å/pix.
x 640 pix.
= 432. Å		0.68 Å/pix.
x 640 pix.
= 432. Å

Surface

Projections

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Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.675 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.15
Minimum - Maximum-0.3870784 - 0.9564988
Average (Standard dev.)0.00051257527 (±0.023445338)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions640640640
Spacing640640640
CellA=B=C: 432.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #2

Fileemd_33528_half_map_1.map
Projections & Slices
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Density Histograms

Histogram

Histogram (log scale)

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Half map: #1

Fileemd_33528_half_map_2.map
Projections & Slices
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Histogram

Histogram (log scale)

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Sample components

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Entire : a membrane protein complex

EntireName: a membrane protein complex
Components
  • Complex: a membrane protein complex
    • Protein or peptide: Ctap3
    • Protein or peptide: Ctap4
    • Protein or peptide: Ctap5
    • Protein or peptide: Toc75
    • Protein or peptide: Toc90
    • Protein or peptide: Tic214
    • Protein or peptide: Protein TIC 20
    • Protein or peptide: Tic15
    • Protein or peptide: Simp3
    • Protein or peptide: Tic100
    • Protein or peptide: Tic56
    • Protein or peptide: Toc34
    • Protein or peptide: Simp2
    • Protein or peptide: Unknown peptide
  • Ligand: Digitonin
  • Ligand: 1,2-DISTEAROYL-MONOGALACTOSYL-DIGLYCERIDE
  • Ligand: HEXADECANE
  • Ligand: DODECANE
  • Ligand: INOSITOL HEXAKISPHOSPHATE
  • Ligand: 1,2-DIPALMITOYL-PHOSPHATIDYL-GLYCEROLE

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Supramolecule #1: a membrane protein complex

SupramoleculeName: a membrane protein complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#14
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 812 KDa

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Macromolecule #1: Ctap3

MacromoleculeName: Ctap3 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 51.982613 KDa
SequenceString: MADGPSPIRI VLWNDGGESL AAGVEDEEQQ QVLHSFADLV GSAIDAVLEL PQFRHVEAVT AEAEEDEPGL SIGFDAGSGD GEVDIDNLK GRLDIAGLLL GSAQLPEELA EVAAVEVTDE EEGTTELQFT DEGLVQQLQA VVKRAKLEKR YNDWVAGVAE S LGPALDAA ...String:
MADGPSPIRI VLWNDGGESL AAGVEDEEQQ QVLHSFADLV GSAIDAVLEL PQFRHVEAVT AEAEEDEPGL SIGFDAGSGD GEVDIDNLK GRLDIAGLLL GSAQLPEELA EVAAVEVTDE EEGTTELQFT DEGLVQQLQA VVKRAKLEKR YNDWVAGVAE S LGPALDAA AGGVEVTEMP VDPYDVLQAV VAQLIRVAGV SPPAPSLFSR TGALVGGVLG APRSAVRQVT KRLGRAQRLW WR LEDVVVD GSKLALRLAV KAARPVLVGF VLHRVLKTLD RSRQLEYRLA RMGPEEAREA YYEAVLGKDW KQQLQADWDK ALE DVDAGL VTDEINHEKR LMTAAQLRRL EVEEWDKQRM KNFYLASFGG LRWFDQMEQA LHNPLFIESR GWTDPVQNWV GQNR TYMDD LPAGQYMAGV GNAAIRIKEA ELKRKLTDVE RAHVLARGGA VAGGLLPQQP TDPATLAVAV GGAFVPSVAG KR

UniProtKB: Flagellar associated protein

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Macromolecule #2: Ctap4

MacromoleculeName: Ctap4 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 39.025082 KDa
SequenceString: MGASQESELD FVPRLSFLPI EWRSIGSAFG LKDKSGAAAN GRATFTVRQG VDAAELTSTG RVIDGQADVG ASLKLNTLAI GVSASNITF HSGLDDPTAA AAQRSSLIPS LKLTAAKQFK RDNYIAVSYD LKHQKPELSA CWTGEAGADR ATLLVNVDPV M RSVKLAAA ...String:
MGASQESELD FVPRLSFLPI EWRSIGSAFG LKDKSGAAAN GRATFTVRQG VDAAELTSTG RVIDGQADVG ASLKLNTLAI GVSASNITF HSGLDDPTAA AAQRSSLIPS LKLTAAKQFK RDNYIAVSYD LKHQKPELSA CWTGEAGADR ATLLVNVDPV M RSVKLAAA VRTPGPEWRK VLYNDETDLL EYPADDGARH TLYVQHEVRG RDLLHATRLG CRLDLGRLVN YVVDFVDYRI EE NIPSFVW NVPLLPQLYS LLVPADNDEQ VRHRITGWEL DVSHDFARSG LLPVVAISKT SKKLLGGGTL TASYDAAARE AGV SLSRKG VSVGARVARA EGAAGGLSAG WGRPSIHVAV EPLGLLQ

UniProtKB: Uncharacterized protein

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Macromolecule #3: Ctap5

MacromoleculeName: Ctap5 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 42.710965 KDa
SequenceString: MQLGQLRQPL RACQDQRLTR GVPLARRQLV VVSNWNPLGG KGGGNAEQPG GEEPIQDELL KLLRGGWVLL SNLALFLVFS SFLHRSLNW FVQTELLVAV GAPQQAGERV VGKFFEAIEW VERNILGWKL PGDEEAEDAT SKVYEVLQNY TPAEAAYSFA Q LKYKDLTH ...String:
MQLGQLRQPL RACQDQRLTR GVPLARRQLV VVSNWNPLGG KGGGNAEQPG GEEPIQDELL KLLRGGWVLL SNLALFLVFS SFLHRSLNW FVQTELLVAV GAPQQAGERV VGKFFEAIEW VERNILGWKL PGDEEAEDAT SKVYEVLQNY TPAEAAYSFA Q LKYKDLTH KERELFHKAY ALRHFERRDG RPGDVDAAEL QAVKDRLDPL EADRRAYAAA KAAGRLDEYW AAPGREATYQ RI VGAPRIA ARQCEMASML KGLQAVLPAM ELLAQLQVAQ FVYAASKASK SRQQDDFKLQ LQTFYGNVLD EQCQLRCMLL NVQ LPMALV TVFVPQYCFC LLDRVVLPRR ECGSASTLGS LLRACGRPGS ACHAGGCSAQ RRDPLTI

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Macromolecule #4: Toc75

MacromoleculeName: Toc75 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 87.333289 KDa
SequenceString: MQGLKATPGL RASSGRPTLR TARTALVVRA HASQPSSSNH AEQQECSSSG SGVSVNQPSA LGRLGKFGLS SALSAFVLVP NFGGFGGNG GNRGGGGGGG GGGGSGGQGQ PDGGLPLPLY ELAEESSDEK EKQQKDDKRN WKNLVTDSED LEEKPGERSG T NRCVEIVI ...String:
MQGLKATPGL RASSGRPTLR TARTALVVRA HASQPSSSNH AEQQECSSSG SGVSVNQPSA LGRLGKFGLS SALSAFVLVP NFGGFGGNG GNRGGGGGGG GGGGSGGQGQ PDGGLPLPLY ELAEESSDEK EKQQKDDKRN WKNLVTDSED LEEKPGERSG T NRCVEIVI EGWPDVGNLP TADELKDLLT VQEGHIFEKQ DLLDDRRKLE IQYEDYIAEV EIRTEYVDGK SNHQRVVYKF TP HQFRGIN AIDIKGAALM PASEVERICN ECLPKQPYMV DIAVMDKVRN RIEQWYQSRG LPFCYVGFFD GMDDGILRAN VTE AKIDNV SVRFVRPKLT GDSELEYSVY DEGKVVKADK IIEASGFQRG HHYHVEDGYD AMNSIFACGL LEDINIEPEQ DPSD VNKIN VKIRCEEVQP KSMELDLDWS FQLKNGIPSI NRQSLIPGGS VEVSHENLFG NSESATLSLS ASDWRNPSAD LGFSV AYSE PFYKPHTTRN AQLFNTRKTS TIFTPGGESE VPPVFVDRFG LKGWTSQITG QDNKVEHALM LQLVSTLDEN GQVVAK GTK VQRGYYADNG PPTTNSGNGR DLSLSYQGFF ALDNVRFING NQLGERMLFQ VDQGLNPSIS LPGGRKLGLS GGIYNRA TA SYTKFLEAPF LPKLTTEQLW KERKAPNTVV LHAKAGNALG DVAAYDYFSL GGPYSVRGYS HGEIGAARRF LELATEVR V PLKNYGLPGT AYGFVEYATD LGSGRELNGN PTEYYRKPGR GMSYGLGLKA LGACRFEYAR DCNAGTGTFL VNFGERF

UniProtKB: Bacterial surface antigen (D15) domain-containing protein

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Macromolecule #5: Toc90

MacromoleculeName: Toc90 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 102.415258 KDa
SequenceString: MGGGLPPKRS EVAESQPASS ASSSAAAPAP AAETAGPKLP PRPAGLGLGG AGLLPSRGAA AAPSAGSATG TPAAAASSSL QQQQQQPAP PATQPAAHAA PAAPAGLGGA GLKPMLPPRP AAAAGAGAAG AAAAKPTPAP APAAAPVPAA APPPRPAMPN P AAGMSLPP ...String:
MGGGLPPKRS EVAESQPASS ASSSAAAPAP AAETAGPKLP PRPAGLGLGG AGLLPSRGAA AAPSAGSATG TPAAAASSSL QQQQQQPAP PATQPAAHAA PAAPAGLGGA GLKPMLPPRP AAAAGAGAAG AAAAKPTPAP APAAAPVPAA APPPRPAMPN P AAGMSLPP RPVVAAAPPV LAAAGSDAVV DPSEDRRVQR VQRIAHDTRV RLIRAASRLG LAPRTDQVAQ FLQAIERSER MV GAQHYKG SRRVDLLAAA EREARLAEER EGAAAEAVAG LRVKILVLGM TGTGKTELIN SLLNRPAGSR TNAFREATRR VRV VRGDHN GIPLTFIDTP GLHASASRTA DNRAILRAVR AAYRWHKPDY VFYVDRLDAT RPGFGEMGLL GLITESLGAG VWRN TMAVL THAHAARTAF GGQYDVNSRQ RRNIVSQLLR QAAGDQQSRN PVFLADCHPA CPTNSLGQPV ILEGPTAVPW KQQLL VQLV GYKSYNVATS AFKDLAKAKA GKAAAGAAGG ARGPQDIFKQ MMRSRLPPMT FFVEQMSEGV LKPEGWATME TVAGLG EEV TEDEGAESFN HVYYRQMYEL AVAGDPWAQR EYAAMLRAYD KGCESYRASY EEADVDANVE YGVESYVVDP IDFGPSF DP EDMYSHRHAY AEAADAGVTV IPSQDYYGPE HDDPLNGIVF QYEAQPFSRH GWGGVPFDLT VCCEKDKTSL CLQGETHV S LVHSVPPFGP RHITQVTGSW EVLRPNIKDV MYQLEVDTFK DGLLGKSDHA GCGLMLARLG EGGDPRKGPT AVGVRLQDT LRVGPFKLEA CASKVAVQGP TGGKEEGWGA RAFVGYDWLP GLGMAFDFIQ ERTPEEGGKR LRGYGANFTY DWEALGAAFG MEVDYVAAS ESVFVSVNAF SGNDYRLGWL LLLPAVNYFK ETVSSLWARL RGAGGGEGEE GEELEEEGEG EEGDDEEAMM M MAQEGDL

UniProtKB: AIG1-type G domain-containing protein

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Macromolecule #6: Tic214

MacromoleculeName: Tic214 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 232.535719 KDa
SequenceString: MITFTFMSLV TSVKDYVEIT HKLIEIEPLK NYTEFGAVFT YFIFSIGEFF KNFFSFSFLN NIWSIPIIIP DIASAMISEV SVLDGYFHN AFTFLETSVN TTTNPSLVIF EKFVIGIINS LFLILPTSTS HLITLRRFVM QGLEAGYMAG LGTLAGNFLW L ASIILGWR ...String:
MITFTFMSLV TSVKDYVEIT HKLIEIEPLK NYTEFGAVFT YFIFSIGEFF KNFFSFSFLN NIWSIPIIIP DIASAMISEV SVLDGYFHN AFTFLETSVN TTTNPSLVIF EKFVIGIINS LFLILPTSTS HLITLRRFVM QGLEAGYMAG LGTLAGNFLW L ASIILGWR FFVIPWLSLD IFRYLLGFVL LVKYIWDSSK ERRMALEDLS KWKIFLLNFL LALTEQSCIY PFISNLSFGP DA SILEGFP VDNYPQFLLI HGAYLLGILF GSFSLLQFTC WFWENPAFSI YLWITTKSSL KISTSSYYKI LNFTFLYATM LCA IASIPY YGLDYTITNP IGLVPQDRIL NQKKSQSDPD KLITETAFLN LNPTDKNSRI RDGVHARRER WKQRLIKYQA FDAS TYDQG VYDFLTIEDL NYGFDRFWLR RKMRNHQIRF RLFPGPWMRS LKKQLNNPAN PSLETSTKAA SGPRVEFFRI LFEQF YHPN FHDRAAMQTN PAEARNKFIS TSPLASTESK KALNSTFSLG NINNSSTGIE GLVLTNTQAT LLPTDLQTKR TIKPGL IYT NSALRKFVRN VNTRLNLKLL NSKETNLTTK YKSQFIYSKR WKSIFSKIQP LQNGTTRKSY QLFRNVAKQI LVTPDAK SL KLITINQKLS LKERKLLELR TQYNNNSTLT TTAPLTLVRP LNVYLQKEEA FKRKLRYYGT MPMRKLTVGN QAPYFKAL M KRGFYYYKPT LRWRKTLYVA SLRRGFRKKS RKQRILVMPS NQQNFNNTLD NTKTNINQNN LANPLGGNEV PMYGADGEN SLITKPTHSY TVLGKRASRY RHQIYKDVLQ HWYYTPFNRL LMKFDVDAFI NRQPKSHFLT KNEERALHIR RFLLSEHYDT LRWYTYMQH YKTMKTNIGG TKSFANRAYN QQFQGTFKKI RHLFAITPKQ GDFYTLKFDQ PLYNDNKLKD NLYFHEELLT D YYNGTNLQ TNQTSNISVN STTTFIDNSL RTTQLPVPSS SFDIVNQSST LIGLTTMQNA LRKNVVESTL TSLNSDGEAA TS QPKLNFV YSELFVKLIK ECKKRIHDQT FLKNYITHRI EKREQLNQEQ TKELNKRLEK LKVWLNSDKG SISKLQNTPV QDP NISSPD KVLTTAMQKA VNESISLSGI MPSDKIKTTY GNLTNAYTIK TENAILTKLN VINQLTNNET TTQKNTLIKS IGVN KIQTV LQTIITNFKS SLYNQTQLLR VKTDKDLQWW RTKQRVITKR KSARKRDRFK KQIAVVNKKL AALSKKVETE KSNLY QTLY GNYEISDYLL RNVPTGSSAV IDSTVLRKKQ DNQAYLPKET NNVQFNSFVD SNNNVWQTFF AKKLRKKISS KGRRYR SLS LARYLTATRK PRLVGLDNLT KIDNITTLQG AFITKEEKQD SLNLTIQRKQ ELTNSLKKSQ IKKRSRHSWK KRSRHQF SR NHYKYRKRHT HGNGKLRVMN KKLKKFKATN ELRQWWWNSF LPRYLSNLQV NNSTLTNKNV SFKPLSNTNS VPSTNMAS P TTSRNLLDNL NSSNQISTSA SMNQNIVTES VKVETNQVYL PEGEKSFDIT SMTTTLPFYA GWDESLKKFV VTNRLLSRR DAGLSVNNNP QEINFTNPPI QGLNEGSFLY WQTEMPFNSY NIDQFITTNQ SFYAPLGWRR FEFRHSILKT WVNNTKAGNN NIKKKTLII SLKNLQPLKS SQQKQNQIKT KKLVARRIKK RYKLLKQMPN QLMYSPTGPL LTEVLPSHYI SVFDQQYRLP R NRYLKRNP LKTLKKTTLL ALMDSSKQTN GVNKEFTLRK RVKPRRKYHR KRFIKKDGLI FPRRTKFNTN TTLTGNALIT NN VNSIEED DLRWRPSSRT KQKRKDNTRS SAASKTKSNK RVKTNPLRLR QLRRREFQQV LKPLQRYIPQ NGGFTWPGDY LRL EIVEMP KLKSINIKKT SLKQKINVQP VGIMPRKYLI EKHNIKVLKK KLSQAYSTQQ LTKVVQEYKN LIQNSPPAI

UniProtKB: Uncharacterized membrane protein ycf78

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Macromolecule #7: Protein TIC 20

MacromoleculeName: Protein TIC 20 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 29.54548 KDa
SequenceString: MSALLSQASG FSGLALGSSA SSKRLCKPLC RRPAVCVQAA HRPASAVSSG VPSTGFGPVS PLQRRPTLPT RKLVVASSQS GGRAEDAEK QDWKFGRNEG PMTWPWKLMC AILYMLPWVD VTEKTVYFVE RFPAFVWTEY FSEPFEHWYN IHEYAPLFIF F ATYLGIVR ...String:
MSALLSQASG FSGLALGSSA SSKRLCKPLC RRPAVCVQAA HRPASAVSSG VPSTGFGPVS PLQRRPTLPT RKLVVASSQS GGRAEDAEK QDWKFGRNEG PMTWPWKLMC AILYMLPWVD VTEKTVYFVE RFPAFVWTEY FSEPFEHWYN IHEYAPLFIF F ATYLGIVR NKKIPHVARY HVMMGVMLDI VAMILIVTEE NLPTGVLWTP WSDLFYALMF WFIFLLVIYC LFFCFLGWYC EI PLISEGV YLQIEQAEQL GQ

UniProtKB: Protein TIC 20

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Macromolecule #8: Tic15

MacromoleculeName: Tic15 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 14.662776 KDa
SequenceString:
MDEEPPFNLA LNVYKGPASI PHASAEVFGA FFLATNTALL AHMFPGKLFG SELHVRKWDP DYLASCCNEQ GMRREALSGK KPNLWLLGG GPRLVNDSWE RMWWNNLHWK RWKVPRTGPA FPQDMYWQ

UniProtKB: Uncharacterized protein

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Macromolecule #9: Simp3

MacromoleculeName: Simp3 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 21.18565 KDa
SequenceString:
MATCAAVSLA TAVFQVAAKY VNKARDAVAS ALPSPSNIDG VQQDWRQQVK ELKKHTRVLG FVLPLAKAFA AESLTSVYAR TFERAAFGF AKMYLFCLFM RVLLSWFPSI DWNSQPWAFL RLITEPYLQI YRGILPPLFG QLDFTPLFGF LILQDVVELM S PVYTLGHA KDTSMFWTTT DIMCYFDGH

UniProtKB: Fanciful K+ uptake-b family transporter

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Macromolecule #10: Tic100

MacromoleculeName: Tic100 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 105.817695 KDa
SequenceString: MASKKGTDAP AALTDPLKED PTVIRDEAQF PEPSLYFKVF ESEAGEPEAK IRADVNKLYD RWIEKYGRRW PEDGINTEDM VWLAEEANK RKRAKPRPRG TVAAEKTEYE DEFMPDPTVG APVSAADAAK AARRAKKDRK KKKAAGGAEQ PAGPRTNYEK T VAGGKWVT ...String:
MASKKGTDAP AALTDPLKED PTVIRDEAQF PEPSLYFKVF ESEAGEPEAK IRADVNKLYD RWIEKYGRRW PEDGINTEDM VWLAEEANK RKRAKPRPRG TVAAEKTEYE DEFMPDPTVG APVSAADAAK AARRAKKDRK KKKAAGGAEQ PAGPRTNYEK T VAGGKWVT DEFESADYEA GNLEKLWDMY LWDREGKPTM MPDTPAAQQE GEESEDFDDF YTAYRPRDVD SEEAREAVWA TD EFESDED NTESEWAPEY VGAGLGLVAE DPLNPQYSLR HSNHPLAPFP GEPLKWASYV YPDFTTFEGL SKQSIPHGMG VMT FGTGTG AGFAMSQTRY GDKYEGEFQA GYAHGLGQFT SEASGEVYIG EFFAGQRHGC GMTLDMKPYF YLLERGVDPV EAYR RTAGA IMKNVEVRTW YRGNKLGDAK EDEVVEINVL KDELDDPFEI ALRNSLHDAK LRKWKAMSPQ DKAMDRIVSI IERVQ RRNP GRFGAYYRED EKGRVRPVLD SDGADTDFDS VDMIQGVDTD GDLGPGWEGA TDSEENPMDP RIRELMAAEG MDDKLE DEG FKDTVLGSAI INPYTGLDMK TYLDGKERHQ AELVSVYKAS REGRKYLNKV RKDKGGAAKD DESSYVEDDA ASGHPGA LL SREAEDDRLA RLYEQAGVSK EDERRVEGLA ARWRRLLAAD EEEVLGGAVG AFRRPGNPLA ANDSDTGFET ESDMMEMC D IPEILGTVQE ARQIVERARM WRFKPYGEVG LRMAQDANGS PVSLMQEPLH YPHGTKFMAP GPLGLCHAVP DDPSLRQEM AKVAHNYAAI YRMYNFDWDP EPGTVQYKID QRIRRAQELR NNAMARYLAA ADEVLRDGAA PAGEGDQALL LASTSTGAPE AFDGQGNAS GSGSSSALSS RGGSMFASMT LSRPAPMAGV VSLGRAARVV LGAFADAAKS VPMARPRLAR PSGRRQ

UniProtKB: Uncharacterized protein

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Macromolecule #11: Tic56

MacromoleculeName: Tic56 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 28.046527 KDa
SequenceString: MSEPGAGPSD GQVVTRKIRL HKVMRPLDES SPSSQEQHMD RRLAEILPAI ADLPVPGPSG AGGSPADARA VEMRRLRGTQ QELAQMEAM ELATLFDMSK PHPLDNAAPA TPWKGELRPV PRKIVLSPYQ YEMINYQRML MRKNIWYYRD RMNVPRGPCP L HVVKEAWV ...String:
MSEPGAGPSD GQVVTRKIRL HKVMRPLDES SPSSQEQHMD RRLAEILPAI ADLPVPGPSG AGGSPADARA VEMRRLRGTQ QELAQMEAM ELATLFDMSK PHPLDNAAPA TPWKGELRPV PRKIVLSPYQ YEMINYQRML MRKNIWYYRD RMNVPRGPCP L HVVKEAWV SGIVDENTLF WGHGLYDWLP AKNIKLLLPM VRTPEVRFAT WIKRTFSLKP SLNRIREQRK EHRDPQEASL QV ELMR

UniProtKB: GYF domain-containing protein

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Macromolecule #12: Toc34

MacromoleculeName: Toc34 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
EC number: Hydrolases; Acting on acid anhydrides; Acting on GTP to facilitate cellular and subcellular movement
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 44.598613 KDa
SequenceString: MAQPPRPAEE YDDDVQEDED ELKEGELDDD ESHEAASEGG EAAAGDEEAE DDEQDEEDGD EDSQPWAGLN RLPERDDMLD ILNELRAEG RKQLTVLLLG KSSVGKSSLI NSLLGEAVVR VQAFKLQADT DITTTVVRQV AVGNSEVDGF RLKLIDTCGL E DPEAGDTV ...String:
MAQPPRPAEE YDDDVQEDED ELKEGELDDD ESHEAASEGG EAAAGDEEAE DDEQDEEDGD EDSQPWAGLN RLPERDDMLD ILNELRAEG RKQLTVLLLG KSSVGKSSLI NSLLGEAVVR VQAFKLQADT DITTTVVRQV AVGNSEVDGF RLKLIDTCGL E DPEAGDTV NLGALSKIAE DVRGVGIDVV LYCDRLDLYR VDPLDKAIID AISSTFGRGI WRRTVVALTH ANLVQTPPGT DY DSFVNGR VRLIRGAVRG PLFFRPSLPV ALVENSETCP VSSESGFRVL PDGEPWLVAL VSQLVDMAAA RRRPYKYHPR LSS KPSHRF RWLLPVAIAA EVLFYRRFLH PRLDDNQRRV EREEERVWAL RGQQRRALGL HRPHRPDKDA AWRLEQMYDD D

UniProtKB: Translocase of chloroplast 34 homolog, chloroplastic

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Macromolecule #13: Simp2

MacromoleculeName: Simp2 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 13.426158 KDa
SequenceString:
MSSDVQAKLS GLLGDIGVKC TLAFAGTVAA GAAIVVPSGK QVEAASLDIY GRPPSQLLPN ERRAAEFAAG HRRWKGFVDN SIYSWTRTL PGHDNPIVNP YKGPRRPQRP QQKLEEEVEA AAKQE

UniProtKB: Uncharacterized protein

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Macromolecule #14: Unknown peptide

MacromoleculeName: Unknown peptide / type: protein_or_peptide / ID: 14
Details: Chain X belongs to an unidentified protein, which contains an alpha helix. The input sequence is a tentative one based on the model. The identity of this chain remains to be revealed in the future.
Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 1.863077 KDa
SequenceString:
AAAAAFAAFA GFAFAAFAAA G

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Macromolecule #15: Digitonin

MacromoleculeName: Digitonin / type: ligand / ID: 15 / Number of copies: 5 / Formula: AJP
Molecular weightTheoretical: 1.229312 KDa
Chemical component information

ChemComp-AJP:
Digitonin / detergent*YM

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Macromolecule #16: 1,2-DISTEAROYL-MONOGALACTOSYL-DIGLYCERIDE

MacromoleculeName: 1,2-DISTEAROYL-MONOGALACTOSYL-DIGLYCERIDE / type: ligand / ID: 16 / Number of copies: 5 / Formula: LMG
Molecular weightTheoretical: 787.158 Da
Chemical component information

ChemComp-LMG:
1,2-DISTEAROYL-MONOGALACTOSYL-DIGLYCERIDE

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Macromolecule #17: HEXADECANE

MacromoleculeName: HEXADECANE / type: ligand / ID: 17 / Number of copies: 1 / Formula: R16
Molecular weightTheoretical: 226.441 Da
Chemical component information

ChemComp-R16:
HEXADECANE

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Macromolecule #18: DODECANE

MacromoleculeName: DODECANE / type: ligand / ID: 18 / Number of copies: 7 / Formula: D12
Molecular weightTheoretical: 170.335 Da
Chemical component information

ChemComp-D12:
DODECANE

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Macromolecule #19: INOSITOL HEXAKISPHOSPHATE

MacromoleculeName: INOSITOL HEXAKISPHOSPHATE / type: ligand / ID: 19 / Number of copies: 1 / Formula: IHP
Molecular weightTheoretical: 660.035 Da
Chemical component information

ChemComp-IHP:
INOSITOL HEXAKISPHOSPHATE

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Macromolecule #20: 1,2-DIPALMITOYL-PHOSPHATIDYL-GLYCEROLE

MacromoleculeName: 1,2-DIPALMITOYL-PHOSPHATIDYL-GLYCEROLE / type: ligand / ID: 20 / Number of copies: 5 / Formula: LHG
Molecular weightTheoretical: 722.97 Da
Chemical component information

ChemComp-LHG:
1,2-DIPALMITOYL-PHOSPHATIDYL-GLYCEROLE / phospholipid*YM

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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

BufferpH: 7.5
Component:
ConcentrationFormulaName
50.0 mMC4H11NO3Tris
150.0 mMNaClSodium chloride
0.06 %C56H92O29Digitonin
GridModel: Quantifoil / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 2 / Pretreatment - Type: PLASMA CLEANING
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K2 QUANTUM (4k x 4k) / Detector mode: SUPER-RESOLUTION / Average electron dose: 50.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: OTHER / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.8 µm / Nominal defocus min: 1.5 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

+
Image processing

Startup modelType of model: NONE
Final reconstructionNumber classes used: 1 / Resolution.type: BY AUTHOR / Resolution: 2.77 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 796731
Initial angle assignmentType: NOT APPLICABLE
Final angle assignmentType: NOT APPLICABLE
FSC plot (resolution estimation)

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Atomic model buiding 1

RefinementSpace: REAL / Protocol: AB INITIO MODEL
Output model

PDB-7xzi:
Cryo-EM structure of TOC-TIC supercomplex from Chlamydomonas reinhardtii

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