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Open data
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Basic information
Entry | Database: PDB / ID: 6zy3 | |||||||||
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Title | Cryo-EM structure of MlaFEDB in complex with phospholipid | |||||||||
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![]() | LIPID TRANSPORT / phospholipid / phospholipid transport / ABC transporter / MlaFEDB / MlaFE / MlaD / MlaE / MlaF / MlaB / outer membrane / Mla transport pathway | |||||||||
Function / homology | ![]() phospholipid transfer activity / intermembrane phospholipid transfer / phospholipid-translocating ATPase complex / phospholipid transport / ATP-binding cassette (ABC) transporter complex / response to antibiotic / DNA damage response / ATP hydrolysis activity / ATP binding / membrane / cytosol Similarity search - Function | |||||||||
Biological species | ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() | |||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.3 Å | |||||||||
![]() | Dong, C.J. / Dong, H.H. | |||||||||
Funding support | ![]() ![]()
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![]() | ![]() Title: Structural insights into outer membrane asymmetry maintenance in Gram-negative bacteria by MlaFEDB. Authors: Xiaodi Tang / Shenghai Chang / Wen Qiao / Qinghua Luo / Yuejia Chen / Zhiying Jia / James Coleman / Ke Zhang / Ting Wang / Zhibo Zhang / Changbin Zhang / Xiaofeng Zhu / Xiawei Wei / ...Authors: Xiaodi Tang / Shenghai Chang / Wen Qiao / Qinghua Luo / Yuejia Chen / Zhiying Jia / James Coleman / Ke Zhang / Ting Wang / Zhibo Zhang / Changbin Zhang / Xiaofeng Zhu / Xiawei Wei / Changjiang Dong / Xing Zhang / Haohao Dong / ![]() ![]() Abstract: The highly asymmetric outer membrane of Gram-negative bacteria functions in the defense against cytotoxic substances, such as antibiotics. The Mla pathway maintains outer membrane lipid asymmetry by ...The highly asymmetric outer membrane of Gram-negative bacteria functions in the defense against cytotoxic substances, such as antibiotics. The Mla pathway maintains outer membrane lipid asymmetry by transporting phospholipids between the inner and outer membranes. It comprises six Mla proteins, MlaFEDBCA, including the ABC transporter MlaFEDB, which functions via an unknown mechanism. Here we determine cryo-EM structures of Escherichia coli MlaFEDB in an apo state and bound to phospholipid, ADP or AMP-PNP to a resolution of 3.3-4.1 Å and establish a proteoliposome-based transport system that includes MlaFEDB, MlaC and MlaA-OmpF to monitor the transport direction of phospholipids. In vitro transport assays and in vivo membrane permeability assays combined with mutagenesis identify functional residues that not only recognize and transport phospholipids but also regulate the activity and structural stability of the MlaFEDB complex. Our results provide mechanistic insights into the Mla pathway, which could aid antimicrobial drug development. | |||||||||
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Structure visualization
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Structure viewer | Molecule: ![]() ![]() |
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PDBx/mmCIF format | ![]() | 353.9 KB | Display | ![]() |
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PDB format | ![]() | 290 KB | Display | ![]() |
PDBx/mmJSON format | ![]() | Tree view | ![]() | |
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-Validation report
Arichive directory | ![]() ![]() | HTTPS FTP |
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-Related structure data
Related structure data | ![]() 11548MC ![]() 6zy2C ![]() 6zy4C ![]() 6zy9C C: citing same article ( M: map data used to model this data |
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Similar structure data |
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Assembly
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Components
#1: Protein | Mass: 19593.133 Da / Num. of mol.: 6 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() ![]() #2: Protein | Mass: 11795.488 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() Gene: mlaB, yrbB, A6581_13415, A6592_14225, A6V01_00625, A8C65_02170, A9P13_02945, A9R57_12600, A9X72_02835, AC067_23675, ACN002_3282, ACN68_12355, ACN81_18445, ACU57_14445, ACU90_11565, AM270_15510, ...Gene: mlaB, yrbB, A6581_13415, A6592_14225, A6V01_00625, A8C65_02170, A9P13_02945, A9R57_12600, A9X72_02835, AC067_23675, ACN002_3282, ACN68_12355, ACN81_18445, ACU57_14445, ACU90_11565, AM270_15510, AM446_04320, AM464_08585, AMK83_06340, AML07_19340, APT94_23200, APZ14_04295, ARC77_12405, AU473_05735, AUQ13_03075, AUS26_18260, AW106_13275, AWB10_02850, AWG90_020620, AWP75_14100, B9M99_10250, B9T59_16250, BANRA_01811, BANRA_03804, BANRA_03999, BANRA_04207, BB545_09780, BE963_06830, BER14_09135, BHJ80_21985, BHS81_19200, BHS87_17970, BIQ87_18040, BIU72_05405, BJJ90_02770, BK292_10220, BK296_13050, BK373_10175, BK375_18740, BK383_15965, BMA87_01295, BMT49_25595, BMT91_11610, BOH76_04915, BON63_14235, BON66_12205, BON69_26555, BON71_25170, BON72_13935, BON75_23145, BON76_05525, BON83_09240, BON86_01565, BON87_18195, BON92_11035, BON94_12335, BON95_22020, BON96_06355, BTQ04_16475, BTQ06_11870, BUE81_19690, BvCms2454_04822, BvCms28BK_03527, BvCms35BK_01543, BvCmsHHP001_01202, BvCmsHHP019_05314, BvCmsKKP061_00888, BvCmsKSNP073_02163, BvCmsKSNP081_00287, BvCmsKSP011_02050, BvCmsKSP024_00283, BvCmsKSP026_00502, BvCmsKSP045_01650, BvCmsKSP067_01464, BvCmsNSNP036_02151, BvCmsNSP006_04640, BvCmsNSP047_04118, BvCmsNSP072_03187, BvCmsOUP014_03632, BvCmsSINP011_04458, BvCmsSIP019_00407, BvCmsSIP044_00137, BVL39_10130, BW690_19360, BWI89_22070, BWP17_10155, BXT93_02550, BZL31_05925, BZL69_07715, C2M16_04215, C5715_22795, C5N07_18070, C5P01_18835, C6669_12020, C7235_03225, C7B02_15875, C7B08_03565, C7B18_13830, C9098_07790, C9114_15455, C9141_00390, C9160_02340, C9162_04110, C9201_13875, C9306_00415, C9E25_11270, C9E67_03375, C9Z03_07490, C9Z28_00385, C9Z29_07190, C9Z37_14660, C9Z39_14275, C9Z43_00380, C9Z68_06245, C9Z69_11175, C9Z70_06765, C9Z78_03315, C9Z89_09065, CA593_10345, CDL37_12215, CF006_16665, CG692_07300, CI641_009255, CI693_03675, CI694_20835, CIG45_16045, CJU63_19140, CJU64_19020, CMR93_08835, CO706_14630, COD30_12915, COD46_10760, CQP61_03650, CR538_03070, CR539_21250, CRD98_04715, CRE06_06830, CRM83_23500, CRX46_03745, CVH05_10780, CWM24_22870, CWS33_08375, D0X26_09090, D2184_10505, D2185_00445, D2188_02580, D3821_08325, D3822_14575, D3O91_07625, D3P01_09160, D3Y67_18290, D4011_13725, D4074_02435, D4628_02655, D4636_02890, D4638_00790, D4660_02830, D4718_09085, D4L91_11950, D4M06_11225, D4U85_14330, D5H35_09780, D5I97_10670, D6004_08125, D6C36_14160, D6D43_15510, D6T60_15390, D6T98_02410, D6W00_14015, D6X36_05360, D6X63_01430, D6X76_04335, D7K33_12855, D7K63_12220, D7K66_04495, D7W70_15855, D7Z75_02945, D8Y65_13475, D9610_07810, D9C99_05120, D9D20_06275, D9D31_14515, D9D43_03615, D9D44_04410, D9E19_10400, D9E34_11740, D9E49_09595, D9F17_07200, D9F32_01240, D9F87_05870, D9G29_07025, D9G48_11290, D9G69_01100, D9G95_17085, D9H53_01195, D9H68_07560, D9I18_07175, D9I88_07240, D9J03_05150, D9J11_01130, D9J44_07850, D9J52_07915, D9J58_06110, D9J60_11185, D9J63_16910, D9J78_05130, D9K02_06080, D9K48_12255, D9L89_05370, D9S45_00255, D9X77_02330, D9X97_03120, D9Z28_03370, DAH18_20610, DAH26_17315, DAH30_12875, DAH32_13975, DAH34_01980, DAH37_04255, DAH43_16780, DBQ99_04040, DD762_13125, DEN86_08060, DEN89_09405, DEN97_16135, DEO04_19605, DEO19_06430, DIV22_02975, DJ503_19040, DK132_07375, DL257_05650, DL292_05795, DL326_06060, DL455_12700, DL479_06590, DL530_10055, DL705_01300, DL800_23275, DLT82_02655, DLU50_08550, DLU67_08375, DLU82_10330, DLW60_07490, DLW88_04795, DLY41_08155, DM102_15165, DM155_07645, DM267_03265, DM280_09055, DM296_04835, DM382_07360, DM820_04625, DM962_02640, DM973_04650, DMI04_04270, DMI53_12715, DMO02_15405, DMY83_09920, DN660_01330, DN700_04595, DN703_01080, DN808_01890, DNB37_13705, DNC98_07160, DND16_06535, DND79_01330, DNI21_03635, DNJ62_12070, DNK12_02835, DNQ45_01620, DNW42_05405, DNX19_05580, DNX30_11365, DOE35_12610, DOS18_13030, DOT81_04255, DOU81_12375, DOY22_14085, DOY56_12245, DOY61_04695, DOY67_01285, DP265_06555, DP277_10105, DQE83_16545, DQE91_01265, DQF36_08635, DQF57_03615, DQF71_07160, DQF72_06460, DQG35_02830, DQO13_01295, DQP61_01770, DRP48_13910, DRW19_01295, DS143_06770, DS721_11090, DS732_23695, DT034_01270, DTL43_04295, DTM10_06985, DTM45_14510, DTZ20_12090, DU309_11135, DU321_08670, DVB38_07395, DXT69_05520, DXT71_00440, DXX80_009590, E0I42_14340, E0K84_17700, E0L12_03915, E2119_05870, E2127_02575, E2128_02495, E2129_05760, E2134_15595, E2135_08080, E2855_04145, E2863_03970, E4K55_06660, E4K61_09280, E5P22_07075, E5P28_12910, E5P37_13135, E5S42_17100, E5S46_15725, E5S47_16890, E5S58_02935, E5S61_19420, EA214_08600, EA223_04825, EA225_07080, EA233_06175, EAI42_05495, EAI52_09890, EAM59_01020, EAN70_10745, EAX79_13230, EB476_09250, EB509_07200, EB510_12405, EB515_07025, EBA84_05830, EBJ06_06135, EBM08_05455, EC1094V2_454, EC3234A_53c00700, EC3426_04342, EC95NR1_02575, ECTO6_00563, ED225_04270, ED307_11785, ED600_09400, ED607_10425, ED611_04280, ED903_04655, ED944_07260, EEA45_06940, EEP23_06300, EF082_03990, EF173_17005, EG075_10110, EG599_06165, EG796_01190, EG808_04845, EGC26_04915, EGU87_04215, EH186_08760, EH412_00995, EHD45_17860, EHD63_05275, EHD79_13055, EHJ36_00995, EHJ66_06090, EHV81_00555, EHV90_06170, EHW09_05600, EHX09_11795, EI021_04065, EI028_15050, EI032_12380, EI041_02000, EIZ86_06785, EIZ93_11790, EJ366_25995, EJC75_15225, EKI52_18235, EL75_0496, EL79_0518, EL80_0509, ELT20_05045, ELU85_06925, ELV05_02275, ELV08_02985, ELV15_01830, ELV28_06345, EPT01_00415, EQ820_11545, EQ823_05050, EQ830_04520, ERL57_13435, ERS085365_00538, ERS085366_00551, ERS085374_01930, ERS085379_00371, ERS085383_00695, ERS085404_02185, ERS085406_02933, ERS085416_00356, ERS139211_02331, ERS150873_02331, ERS150876_01029, EST51_02745, EVY14_07725, EXM29_19995, ExPECSC019_00712, ExPECSC038_02702, EXX71_13400, EXX78_16730, EYD11_02645, EYX82_02455, EYY34_10615, F1E19_20375, F7F00_12170, F7F11_09035, F7F18_01205, F7F23_11445, F7F26_00405, F7F29_05115, F7G01_13980, F7G03_16150, F9Z74_11630, FAF34_018885, FE846_13735, FKO60_06240, FNJ69_14865, FNJ83_20565, FORC82_0569, FQ022_08190, FQ915_12490, FQR64_03035, FQZ46_16130, FTV92_21230, FV293_08375, FVB16_04440, FWK02_19645, FY127_07380, FZ043_22035, GHR40_07595, GII67_12750, GIY13_09305, GIY19_08335, GJ11_20810, GJD97_02885, GKE15_04665, GKE22_12205, GKE24_04655, GKE26_04130, GKE29_02675, GKE31_04620, GKE39_10535, GKE46_11210, GKE58_04575, GKE60_04100, GKE64_10535, GKE77_05005, GKE87_00910, GKE93_06185, GKF00_03235, GKF03_05520, GKF28_12045, GKF34_04640, GKF47_04635, GKF74_09220, GKF86_04260, GKF89_14375, GKG12_08325, GN312_06640, GNZ00_05080, GNZ02_01290, GNZ03_06110, GP654_17710, GP661_13045, GP664_12080, GP666_12960, GP689_13875, GP712_11385, GP935_13340, GP946_08670, GP950_18225, GQE30_02570, GQE34_13735, GQE51_04670, GQE64_09725, GQE68_06200, GQE88_20265, GQE93_14850, GQM06_05785, GQM09_11065, GQM13_09420, GQM17_12885, GQN16_09500, GRW42_15935, GRW80_12045, HmCmsJML074_04878, HmCmsJML079_04444, HmCmsJML204_02250, HMPREF3040_00351, HW43_20915, MJ49_14345, NCTC10082_02946, NCTC10090_03637, NCTC10418_00837, NCTC10764_03982, NCTC10766_02903, NCTC10865_00786, NCTC10963_00565, NCTC10974_00682, NCTC11022_03358, NCTC11341_01768, NCTC12650_00838, NCTC12950_00597, NCTC13127_00906, NCTC13216_01244, NCTC7922_03906, NCTC7927_00668, NCTC8179_06070, NCTC8500_00486, NCTC8622_00353, NCTC8960_03188, NCTC8985_05122, NCTC9007_04416, NCTC9044_01237, NCTC9050_03718, NCTC9055_02489, NCTC9062_02576, NCTC9077_00732, NCTC9111_00974, NCTC9117_00877, NCTC9701_00685, NCTC9702_00649, NCTC9703_05076, NCTC9706_02844, NCTC9777_02009, PGD_03924, RG28_21075, RK56_014610, RX35_04279, SAMEA3472043_01825, SAMEA3472047_01897, SAMEA3472055_00376, SAMEA3472070_01148, SAMEA3472080_00969, SAMEA3472090_00572, SAMEA3472110_01519, SAMEA3472112_01503, SAMEA3472114_00459, SAMEA3472147_01902, SAMEA3484427_00584, SAMEA3484429_00758, SAMEA3484434_03617, SAMEA3752372_01803, SAMEA3752553_01163, SAMEA3752557_01851, SAMEA3752559_03249, SAMEA3752620_02365, SAMEA3753064_02960, SAMEA3753164_02324, SAMEA3753290_00124, SAMEA3753300_02107, SK85_03504, UC41_09935, UN86_15380, WQ89_06640, WR15_24440, YDC107_1864 Production host: ![]() ![]() #3: Protein | Mass: 29128.801 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() ![]() #4: Protein | Mass: 27885.162 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() ![]() #5: Chemical | ChemComp-PEE / | Has ligand of interest | Y | |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
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Source (recombinant) |
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Buffer solution | pH: 7.8 / Details: 20 mM Tris-Cl, pH 7.8, 150 mM NaCl and 0.05% LMNG | ||||||||||||||||||||||||||||||||||||
Buffer component |
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Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | ||||||||||||||||||||||||||||||||||||
Vitrification | Cryogen name: ETHANE / Humidity: 95 % / Chamber temperature: 298 K |
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Electron microscopy imaging
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: ![]() |
Electron lens | Mode: BRIGHT FIELD |
Image recording | Electron dose: 40 e/Å2 / Film or detector model: GATAN K2 SUMMIT (4k x 4k) |
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Processing
Software | Name: PHENIX / Version: 1.14_3260: / Classification: refinement | |||||||||||||||||||||
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EM software |
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CTF correction | Type: PHASE FLIPPING ONLY | |||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | |||||||||||||||||||||
3D reconstruction | Resolution: 3.3 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 97672 / Algorithm: FOURIER SPACE / Num. of class averages: 2 / Symmetry type: POINT | |||||||||||||||||||||
Atomic model building | B value: 77 / Protocol: AB INITIO MODEL / Space: RECIPROCAL |