2019-12-25RELPDBePDBePDBe2019-08-052019-09-112019-10-092019-12-25Spanish Ministry of Science, Innovation, and UniversitiesSAF2017-82632-P o-funded by the European Regional Development FundSpainEuropean Regional Development FundP2018/NMT4443 co-funded by Autonomous Region of MadridSpainEuropean Regional Development FundY2018/BIO4747 co-funded by the Autonomous Region of MadridSpainBavarian State Ministry for Education, Culture, Science and ArtsN-BM-2013-246GermanyEuropean UnionHorizon 2020, iNEXT (PID2907) , Grant number 653706SpainStructure of the ESX-3 core complexRivera-Calzada AFamelis NGeibel SLlorca OFamelis NRivera-Calzada ADegliesposti GWingender MMietrach NSkehel JMFernandez-Leiro RBottcher BSchlosser ALlorca OGeibel SArchitecture of the mycobacterial type VII secretion system.NatureUK576321325201931597161doi:10.1038/s41586-019-1633-11476-46870006NATUASEMD-10186associated EM volumeStructure of the ESX-3 core complex6sgwFULLOVERLAPESX-3 core complexESX-3 core complex01234The sample consists of four protein components, EccB3:EccC3:EccD3:EccE3 in a 1:1:2:1 stoichiometry
Molecular weight of the complex without the amphipol micelle: 0.65 MDaMycolicibacterium smegmatis MC2 155Mycolicibacterium smegmatis MC2 1550.65ESX-3 secretion system ATPase EccB3Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)ATCC 700084 / mc(2)1550.0094078832Mycolicibacterium smegmatis MC2 155LEVODRRSFSSRTPVNENPDGVQYRRGFVTRHQVSGWRFVMRRIASGVALHDTRMLVDPLRTQSRAVLTGALILVTGLVGCFIF
SLF3.6.-.-ESX-3 secretion system protein EccD3Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)ATCC 700084 / mc(2)1550.0471141254Mycolicibacterium smegmatis MC2 155LEVOPIVRVAVLAAGDDGGRLTEMALPSELPLREILPAVQRIVQPARENDGAADPAAAPNPVRLSLAPIGGAPFSLDATLDTVG
VVDGDLLALQAVPSGPPAPRIVEDIADAAVIFSEARRRQWGPTHIARGAALALIGLILVGTGLSVAHRVITGDLLGQFIV
SGIALATVIAALAVRNRSAVLATSLAVTALVPVAAAFALGVPGDFGAPNVLLAAAGVAAWSLISMAGSPDDRGIAVFTAT
AVTGVGVLLVAGAASLWVISSDVIGCALVLLGLIVTVQAAQLSAMWARFPLPVIPAPGDPTPAARPLSVLADLPRRVRVS
QAHQTGVIAAGVLLGVAGSVALVSSANASPWAWYIVVAAAAGAALRARVWDSAACKAWLLGHSYLLAVALLVAFVIGDRY
QAALWALAALAVLVLVWIVAALNPKIASPDTYSLPMRRMVGFLATGLDASLIPVMALLVGLFSLVESX-3 secretion system protein EccC3Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)ATCC 700084 / mc(2)1550.0446530782Mycolicibacterium smegmatis MC2 155LEVOSRLIFEHQRRLTPPTTRKGTITIEPPPQLPRVVPPSLLRRVLPFLIVILIVGMIVALFATGMRLISPTMLFFPFVLLLAA
TALYRGGDNKMRTEEVDAERADYLRYLSVVRDNVRAHAAEQRAALEWSHPEPEVLATIPGTRRQWERDPRDRDFLVLRAG
RHDVPLDAALKVKDTADEIDLEPVAHSALRGLLDVQRTVRDAPTGLDVAKLARITVIGEADEARAAIRAWIAQAVTWHDP
TMLGVALAAPDLESGDWSWLKWLPHVDVPNEADGVGPARYLTTSTAELRERLAPALADRPLFPAESGAALKHLLVVLDDP
DADPDDIARKPGLTGVTVIHRTTELPNREQYPDPERPILRVADGRIERWQVGGWQPCVDVADAMSAAEAAHIARRLSRWD
SESX-3 secretion system protein EccE3Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)ATCC 700084 / mc(2)1550.0308133552Mycolicibacterium smegmatis MC2 155LEVOMTARIALASLFVVAAVLAQPWQTTTQRWVLGVSIAAVIVLLAWWKGMFLTTRIGRALAMVRRNRAEDTVETDAHRATVVL
RVDPAAPAQLPVVVGYLDRYGITCDKVRITHRDAGGTRRSWISLTVDAVDNLAALQARSARIPLQDTTEVVGRRLADHLR
EQGWTVTVVEGVDTPLPVSGKETWRGVADDAGVVAAYRVKVDDRLDEVLAEIGHLPAEETWTALEFTGSPAEPLLTVCAA
VRTSDRPAAKAPLAGLTPARGRHRPALAALNPLSTERLDGTAVPLsingleParticleparticle0.3830 mM Hepes pH 8.0, 150 mM NaClETHANE100278FEI VITROBOT MARK IVBlotting time: 3s
Blotting force: -10. bound to Amphipol A8-35FEI TITAN KRIOSSPOT SCANBRIGHT FIELDFIELD EMISSION GUN3002.71.62.675000.0FEI TITAN KRIOS AUTOGRID HOLDERNITROGENFEI FALCON III (4k x 4k)COUNTING1190350.0Images acquired as 55 frames movies at a calibrated magnification of 1.0635 Angs/px1Images were collected in counting mode2066007GctfC13.8FSC 0.143 CUT-OFFRELION126308MAXIMUM LIKELIHOODRELIONMAXIMUM LIKELIHOODRELIONAB INITIO MODELMap to model correlation coefficientREAL76.45