2024-03-06RELRCSBRCSBRCSB2019-12-192020-01-292020-01-292024-03-06Other privateUnited StatesStructure of the bovine BBSome:ARL6:GTP complexSingh SKGui MCilia, ciliopathy, complex, membrane-protein transport, PROTEIN TRANSPORTSingh SKGui MKoh FYip MCBrown AStructure and activation mechanism of the BBSome membrane protein trafficking complex.ElifeUS9202031939736doi:10.7554/eLife.533222050-084X6vbvFULLOVERLAPBovine BBSome:ARL6:GTP complexBovine BBSome:ARL6:GTP complex0123456789Native BBSome complex isolated from bovine retina and incubated with recombinant bovine ARL6 in the presence of GTP.0.5BBSome complex112345678Bos taurusEyeRetinaADP-ribosylation factor-like protein 619Bos taurusBardet-Biedl syndrome 18 proteinBos taurusEyeRetina0.0080705021LEVOMAETKSMFREVLPKQGQLYVEDITTMVLCKPKLLPLKSLTLEKLEKMQQAAQDTIHQQEMTEKEQKITHG3N2W1BBS1 domain-containing proteinBos taurusEyeRetina0.0649391411LEVOMAATSSSDSDGGKGESEANSKWLDSLSDSMANIHTFSACLALADFHGDGEYKLAMGDLGPDGRQPRLKVLKGHTLVSQKP
LPDLPAAAVTFLMASHEPRTPALAIASGPCVYVYKNLKPYFKFSLPSLPTNPLEQDLWNQAKEDQIDPLTLKEMLEGIRE
KAEVPLSVQSLRFLPLELSEMEAFVNQHKSKSIRRQTVITTMTTLKKNLADEDAVSCLVLGTENKELLVLDPEAFTILAK
MSLPSVPAFLEASGQFDVEFRLAAACRNGSIYILRRDSKRPKYCIELGAQPVGLVGVHKVLVVGSNQDSLHGFTYKGKRL
WTVQMPAAILAMNLLEQHSRGLQAVMAALANEEVRIYHDKVLLNVIRTPEAVTSLCFGRYGREDNTLIMTTLGGGLIIKI
LKRTAVFAEGGGEAGPPPSQAIKLNVPRKTRLYVDQTLREREAGTAMHRTFQADLYLLRLRAARAYVQALESSLSPVSLT
AREPLKLHAVVQGLGPTFKLTLHLQNTSTARPILGLVVCFLYNEVLYALPRAFFKVPLLVPGLNYPLETFVKSLSDKGIS
DIIKVLVLREGQSTPLLSAHINMPMSEGLAADE1BN34Bardet-Biedl syndrome 2 protein homologBos taurusEyeRetina0.079911483999999991LEVOMLQPVFTLKLRHKISPRMVAVGRYDGTHPCLAAATQAGKVFIHNPHSRSQHLGAPRVLQSPLESDVSLLNINQTVSCLTA
GVLNPELGYDALLVGTQTNLLAYDVYNNSDLFYREVADGASAIVLGTLGDITSPLAIIGGNCALQGFNHEGNDLFWTVTG
DNVHSLALCDFDGDGKKELLVGSEDFDIRVFKEDEIVAEMSETEIITSLCPMYGSRFGYALSNGTVGVYDKTARYWRIKS
KNQAMSIHAFDLNSDGVCELITGWSNGKVDARSDRTGEVIFKDNFSSAIAGVVEGDYRMEGCQQLICCSVDGEIRGYLPG
TAEMRGNLMDISVEQDLIRELSQKKQNLLLELRNYEENAKAELSSPLNEADGHRGVIPANTKHHTALSVSLGSEAQAAHA
ELCISTSNDTIIRAVLIFAEGVFAGESHVVHPSVHHLSSSVRIPITPPKDIPVDLHLKTFVGYRSSTQFHVFELTRQLPR
FSMYALTSPDPASEPLSYVNFIIAERAQRVVMWLNQNFLLPEDTNIQNAPFQVCFTSLRNGGQLYIKIKLSGEITVNTDD
IDLAGDIIQSMASFFAIEDLQVEADFPVYFEELRKVLVKVDEYHSVHQKLSADMADNSNLIRSLLVQAEDARLMRDMKTM
KNRYKELYDLNKDLLNGYKIRCNNHTELLGSLKAVNQAIQRAGHLRVGKPKNQVITACRDAIRSNNINMLFRIMRVGTAS
SQ32L13Bardet-Biedl syndrome 4 protein homologBos taurusEyeRetina0.0582891331LEVOMAEEKLSARTQLPVSAESQKPVLKKAPEFPILEKQNWLIHLYYIQKDYEACKAVIKEQLQETHGLCEYAIYVQALIFRLE
GNIQESLRLFQMCAFLSPQCADNLKQVARSLFLLGKHKAAIEVYNEAAKLNQKDWEICHNLGVCYIYLKQFDKAQDQLHN
ALHLNRHDLTYIMLGKIFLLKGDLDKAIEIYKKAVEFSPENTELLTTLGLLYLQLGIYQKAFEHLGNTLTYDPTNYKAIL
AAGSMMQTHGDFDVALTKYKVVACAVIESPPLWNNIGMCFFGKKKYVAAISCLKRANYLAPLDWKILYNLGLVHLTMQQY
ASAFHFLSAAINFQPKMGELYMLLAVALTNLEDSENAKRAYEEAVRLDKCNPLVNLNYAVLLYNQGEKRDALAQYQEMEK
KVNLLKYSSSLEFDPEMVEVAQKLGAALQVGEALVWTKPVKDPKSKHQTASTSKAAGFQQPLGSNQALGQAMSSAATCRK
LSSGAGGTSQLTKPPSLPLEPEPTVEAQPTEASAQTREKQ1JQ97Bardet-Biedl syndrome 5 protein homologBos taurusEyeRetina0.038880983999999991LEVOMSVLDALWEDRDVRFDVSSQQMKTRPGEVLIDCLDSVEDTKGNNGDRGRLLVTNLRIVWHSLALPRVNLSIGYNCILNIT
TRTANSKLRGQTEALYVLTKCNSTRFEFIFTNLVPGSPRLYTSLIAVHRAYETSKMYRDFKLRSALIQNKQLRLLPQENV
YNKINGVWNLSSDQGNLGTFFITNVRIVWHANMNDSFNVSIPYLQIRSVKIRDSKFGLALVIESSQQSGGYVLGFKIDPV
EKLQESVKEINSLHKVYSANPIFGVDYEMEEKPQPLEALTVKQIQDDVEIDSDDHTDAFVAYFADGNKQQDREPVFSEEL
GLAIEKLKDGFTLQGLWEVMNA6QLF9Bardet-Biedl syndrome 7 protein homologBos taurusEyeRetina0.0804713751LEVOMDLNLNRADYLQVGVTSQKTMKLLPASKHRATQKVVVGDHDGIVMCFGMKKGEAVTVFKTLPGQKIARLELGGALNTPQE
KIFIAAGSEIRGFTKRGKQFLSFETNLTESIKAMHISGSDLFLSASYIYNHYCDCKDQHYYLSGDKINDVICLPVERLLR
EVPVLACQDRVLRVLQGSDVTYEIEVPGPPTVLALHNGNGGDSGEDLLFGTSDGKLGLIQITTSKPIHKWEIRNEKKRGG
ILCVDSFDIVGDGVKDLLVGRDDGMVEVYGFDNANEPVLRFDHTLSESVTSIQGGCVGKDGYDEIVVSTYSGWITGLTTE
PVHKESGPGEELKFNQEMQNKISSLRSELEQLQYKVLQEREKYQQSSQSSKAKSAVPSFSVNDKFTLNKDDASYSLILEV
QTAIDNVLIQSDVPIDLLDVDKNSAVVSFSSCDSESNDNFLLATYRCQANTTRLELKIRSIEGQYGTLQAYVTPRIQPKT
CQVRQYHIKPLSLHQRTHFIDHDRPMNTLTLTGQFSFSELHSWVVFCMPEVPEKPPAGECVTFYFQNTFLDTQLESTYRK
GEGVFKSDNISTISILKDVLSKEATKRKINLNISYEINEVSVKHTLKLIHPKLEYQLLLAKKVQLIDALKELQVHEGNTN
FLIPEYRCILEEADHLQEEYKKQPAHLERLYGMITDLFIDKFKFKGTNVKTKVPLLLEILDSYDQNALIAFFDAAF1MB52Tetratricopeptide repeat domain 8Bos taurusEyeRetina0.0566864061LEVOMEPLLLAWSYFRRRRFQLCADLCTQMLEKSPCDQAAWILKARALTEMVYVDEIDVDEEGIAEMILDENAIAQVPRPGTSL
KLPGTNQTGGPSPAVRPVTQAGRPITGFLRPSTQSGRPGTIEQAIKTPRTAYTARPIASSSGRFVRLGTASMLTSPDGPF
INLSRLNLAKYAQKPKLAKALFEYIFHHENDVKTALDLAALSTEHSQYKDWWWKVQIGKCYYRLGLYREAEKQFKSALKQ
QEMVDTFLYLAKVYISLDQPLTALNLFKQGLDKFPGEVTLLCGIARIYEEMNNISSATEYYKEVLKQDNTHVEAIACIGS
NHFYTDQPEVALRFYRRLLQMGVYNCQLFNNLGLCCFYAQQYDMTLTSFERALSLAENEEEVADVWYNLGHVAVGTGDTN
LAHQCFRLALVSNNQHAEAYNNLAVLEMRRGHVEQAKALLQTASSLAPHMYEPHFNFATISDKIGDLQRSYAAAKKSEAA
FPDHVDTQHLIKQLEQHFAMLF1N4X0Bardet-Biedl syndrome 9Bos taurusEyeRetina0.0992309141LEVOMSLFKARDWWSTVLGDKEEFDQGCLCLADVDNTGNGQDKIIVGSFMGYLRIFNPHPVKTGDGAQAEDLLLEVHLRDPILQ
VEVGKFVSGTEMLHLAVLHSRKLCVYSVSGTLGNVEHGNQYQIKLMYEHNLQRTACNMTYGSFGGVKGRDLICIQSVDGM
LMVFEQESYAFGRFLPGSLLPGPLAYSSRTDSFITVSSCHQVESYKYQVLAFATDADKRQETEQQKHGSGKRLVVDWTLN
IGEQAIDICIVSFIQSASSVFVLGERNFFCLKDNGQIQFMKKLDYSPSCFLPYCSVSEGTINTLIGNHNNMLHIYQDVTL
KWATQLPHVPVAVRVGCLHDLKGVIVTLSDDGHLQCSYLGTDPSLFQAPKVESRELNYDELDMELKELQKVIKNVNKSQD
VWPLTEREDDLKVSAMVSPNFDSVSQATDVEVGADLVPSVTVKVTLKNRVALQKIKLSIYVQPPLVLTGDQFTFEFMAPE
MTRTVGFSVYLKGSYSPPELEGNAVVSYSRPTERNPDGIPRVSQCKFRLPLKLVCLPGQPSKTASHKLTIDTNKSPVSLL
SLFPGFAKQSEDDQVNVMGFRFLGGSQVTLLASKTSQRYRIQSEQFEDLWLITNELIIRLQEYFEKQGIKDFTCSFSGSV
PLEEYFELIDHHFELRINGEKLEELLSERAVQFRAIQRRLLTRFKDKTPAPLQHLDTLLDGTYKQVIALADAVEENQDNL
FQSFTRLKSATHLVILLIGLWQKLSADQIAILEAAFLPLQQDTQELGWEETVDAALSHLLKTCLSKSSKEQALNLNSQLG
IPKDTSQLKKHITLFCDRLAKGGRLCLSTDAAAPQTMVMPGGCATIPESDLEGRSIDQDSSELFTNHKHLMVETPVPEVS
PLQGVTEE1BHJ5ADP-ribosylation factor-like protein 6Bos taurus0.0210864861Escherichia coli BL21(DE3)LEVOMGLLDRLSGLLGLKKKEVHVLCLGLDNSGKTTIINKLKPSNAQSQDIVPTIGFSIQKFKSSSLSFTVFDMSGQGRYRNLW
EHYYKEGQAIIFVIDSSDKLRMVVAKEELRTLLNHPDIKHRRIPILFFANKMDLRDALTSVKVSQLLCLEDIKDKPWHIC
ASDAIKGEGLQEGVDWLQDQIQSVKTQ0IIM2CALCIUM ION4.0078e-052CAGUANOSINE-5'-TRIPHOSPHATE0.00052317999999999991GTPsingleParticleparticle0.77.520.0Hepes220.0NaCl5.0MgCl24.0beta-mercaptoethanol1.0GTPCARBONHOLEYunspecifiedETHANE100293FEI VITROBOT MARK IIGrids were blotted for 2 s with a -2 offset.. FEI TITAN KRIOSFLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN30050.02.71.12.481000.0FEI TITAN KRIOS AUTOGRID HOLDERNITROGENGIF Bioquantum25GATAN K3 BIOQUANTUM (6k x 4k)294084.056.01An initial map for the BBSome was generated using RELION's implementation of the stochastic gradient descent algorithm using default parameters and a mask diameter of 280 Angstrom.C1BACK PROJECTION3.5FSC 0.143 CUT-OFFRELION3.075201MAXIMUM LIKELIHOODRELION3.0MAXIMUM LIKELIHOODRELION3.08RELION3.0OTHERDuring refinement, the resolution limit was set to 3.5 Angstrom. Secondary structure, Ramachandran and rotamer restraints were applied during refinement.Correlation coefficientREAL43.600000000000001emd_21145_msk_2.mapemd_21145_msk_3.mapemd_21145_msk_4.mapemd_21145_msk_1.mapemd_21145_additional_3.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
320
320320
0
00320320320339.19998339.19998339.1999890.090.090.0XYZ-0.070189060.12762009-0.000117465580.00391617231.061.061.06Post-processed multibody map for the BBSome bodyemd_21145_additional_2.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
320
320320
0
00320320320339.19998339.19998339.1999890.090.090.0XYZ-0.0134972230.127620090.00136945680.00322519691.061.061.06BBSome:ARL6:GTP complex. Chimeric map following multibody refinement.emd_21145_additional_5.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
320
320320
0
00320320320339.19998339.19998339.1999890.090.090.0XYZ-0.044544760.07645348-0.000197145210.0017211441.061.061.06Post-processed multibody map for the BBS1-ARL6 interaction.emd_21145_additional_4.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
320
320320
0
00320320320339.19998339.19998339.1999890.090.090.0XYZ-0.061603580.10264412-0.000167802650.00236216351.061.061.06Post-processed multibody map for the BBSome heademd_21145_additional_1.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
320
320320
0
00320320320339.19998339.19998339.1999890.090.090.0XYZ-0.0241231360.058271375-0.000069948750.00185908521.061.061.06BBSome:ARL6:GTP complex. Unfiltered mapemd_21145_half_map_2.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
320
320320
0
00320320320339.19998339.19998339.1999890.090.090.0XYZ-0.0543354750.09589056-0.0000714392440.00502918571.061.061.06BBSome:ARL6:GTP complex. Half map 2emd_21145_half_map_1.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)