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- PDB-6z12: Salmonella AcrB solubilised in the SMA copolymer -

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Basic information

Entry
Database: PDB / ID: 6z12
TitleSalmonella AcrB solubilised in the SMA copolymer
ComponentsEfflux pump membrane transporter
KeywordsMEMBRANE PROTEIN / AcrB
Function / homology
Function and homology information


xenobiotic transport / efflux transmembrane transporter activity / membrane => GO:0016020 / plasma membrane
Similarity search - Function
Hydrophobe/amphiphile efflux-1 HAE1 / Acriflavin resistance protein / Multidrug efflux transporter AcrB TolC docking domain, DN/DC subdomains / AcrB/AcrD/AcrF family
Similarity search - Domain/homology
Efflux pump membrane transporter / Efflux pump membrane transporter
Similarity search - Component
Biological speciesSalmonella enterica I (bacteria)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 4.6 Å
AuthorsMuench, s.p. / Johnson, R.M.
Funding support United Kingdom, 3items
OrganizationGrant numberCountry
Wellcome Trust109158/B/15/Z United Kingdom
Biotechnology and Biological Sciences Research Council (BBSRC)BB/N002776/1 United Kingdom
Wellcome Trust108372/A/15/Z United Kingdom
CitationJournal: Microorganisms / Year: 2020
Title: Cryo-EM Structure and Molecular Dynamics Analysis of the Fluoroquinolone Resistant Mutant of the AcrB Transporter from .
Authors: Rachel M Johnson / Chiara Fais / Mayuriben Parmar / Harish Cheruvara / Robert L Marshall / Sophie J Hesketh / Matthew C Feasey / Paolo Ruggerone / Attilio V Vargiu / Vincent L G Postis / ...Authors: Rachel M Johnson / Chiara Fais / Mayuriben Parmar / Harish Cheruvara / Robert L Marshall / Sophie J Hesketh / Matthew C Feasey / Paolo Ruggerone / Attilio V Vargiu / Vincent L G Postis / Stephen P Muench / Vassiliy N Bavro /
Abstract: is an important genus of Gram-negative pathogens, treatment of which has become problematic due to increases in antimicrobial resistance. This is partly attributable to the overexpression of ... is an important genus of Gram-negative pathogens, treatment of which has become problematic due to increases in antimicrobial resistance. This is partly attributable to the overexpression of tripartite efflux pumps, particularly the constitutively expressed AcrAB-TolC. Despite its clinical importance, the structure of the AcrB transporter remained unknown to-date, with much of our structural understanding coming from the orthologue. Here, by taking advantage of the styrene maleic acid (SMA) technology to isolate membrane proteins with closely associated lipids, we report the very first experimental structure of AcrB transporter. Furthermore, this novel structure provides additional insight into mechanisms of drug efflux as it bears the mutation (G288D), originating from a clinical isolate of Typhimurium presenting an increased resistance to fluoroquinolones. Experimental data are complemented by state-of-the-art molecular dynamics (MD) simulations on both the wild type and G288D variant of AcrB. Together, these reveal several important differences with respect to the protein, providing insights into the role of the G288D mutation in increasing drug efflux and extending our understanding of the mechanisms underlying antibiotic resistance.
History
DepositionMay 11, 2020Deposition site: PDBE / Processing site: PDBE
Revision 1.0Jul 22, 2020Provider: repository / Type: Initial release

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Structure visualization

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Assembly

Deposited unit
A: Efflux pump membrane transporter
B: Efflux pump membrane transporter
C: Efflux pump membrane transporter


Theoretical massNumber of molelcules
Total (without water)341,2783
Polymers341,2783
Non-polymers00
Water0
1
A: Efflux pump membrane transporter


Theoretical massNumber of molelcules
Total (without water)113,7591
Polymers113,7591
Non-polymers00
Water0
TypeNameSymmetry operationNumber
identity operation1_5551
2
B: Efflux pump membrane transporter


Theoretical massNumber of molelcules
Total (without water)113,7591
Polymers113,7591
Non-polymers00
Water0
TypeNameSymmetry operationNumber
identity operation1_5551
3
C: Efflux pump membrane transporter


Theoretical massNumber of molelcules
Total (without water)113,7591
Polymers113,7591
Non-polymers00
Water0
TypeNameSymmetry operationNumber
identity operation1_5551

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Components

#1: Protein Efflux pump membrane transporter


Mass: 113759.227 Da / Num. of mol.: 3
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Salmonella enterica I (bacteria)
Gene: acrB, A4O23_10955, A4O45_03810, A5895_15265, AAC70_12525, AAL03_14960, AAM89_07765, AAV76_19425, ABK55_05725, ABO08_03475, AC391_10965, ACH21_19020, ACK40_00105, ACK79_15085, ACS18_20650, ACV85_ ...Gene: acrB, A4O23_10955, A4O45_03810, A5895_15265, AAC70_12525, AAL03_14960, AAM89_07765, AAV76_19425, ABK55_05725, ABO08_03475, AC391_10965, ACH21_19020, ACK40_00105, ACK79_15085, ACS18_20650, ACV85_05805, ADQ53_03685, ADQ64_15580, ADT85_17470, AF726_08820, AG46_12710, AG53_15950, AGQ41_04335, AGQ90_03080, AGR09_08325, AGR46_16415, AH060_16145, AH079_04350, AH254_22600, AH360_16970, AH543_16315, AH557_09130, AH612_19020, AH874_13175, AHN36_15795, AHN91_21865, AHQ94_20600, AHS94_22030, AHU36_11045, AHU48_23190, AHX56_08240, AHY46_22370, AHZ72_05105, AIA22_12165, AIB97_16815, AID13_02815, AIE18_09140, AIF85_07160, AL864_02820, AL929_23130, ALX58_14045, AU885_11650, AVA95_03925, AVB28_08510, AVG51_11480, AVH38_04545, AXK79_19705, AXM38_07800, AXM50_14470, AXM56_16685, AXM65_18530, AXM93_08445, AXN14_02155, AXN15_07580, AXN21_06965, AXN39_11550, AXN51_12700, AXN72_08000, AXO05_08570, AXO78_17315, AXO93_15340, AXQ08_14655, AXR02_08970, AXR82_02315, AXR91_01290, AXS04_01695, AXS89_02055, AXT07_05965, AXT19_05915, B9C63_11810, BXE81_09065, BXG02_07180, BXG42_09370, BXH36_10215, BXH66_08040, BXL85_11765, BXO48_06645, BZ557_12735, BZ723_04100, BZ903_08940, BZV91_08875, BZW24_03920, BZW54_04755, C9E93_06140, C9E94_19180, C9F01_08965, C9F05_01290, C9P23_10335, C9Y45_03940, CB317_01000, CC394_01545, CC530_00775, CC828_04535, CE94_19610, CF93_04280, CPY04_22475, CQE41_07070, CQF73_09855, CQH69_04135, CQM91_23105, CSM01_01615, CSN39_10900, CSN48_19880, CSN51_17860, CSO96_14135, CSP66_16145, CST74_11845, CST78_12945, CV491_04375, CWG53_02155, CWG97_11285, D4346_01325, D4392_10020, D4396_04285, D4408_06485, D4427_06950, D4428_05215, D4444_11555, D4471_08450, D4490_01015, D4494_10470, D4507_05270, D4514_03660, D4515_01545, D4546_09390, D4551_09260, D4D97_05485, D4D98_08570, D4E01_09290, D4E37_05350, D4E64_10565, D4E75_00440, D4E90_05185, D4F17_01060, D4F19_04440, D4F20_07155, D4F29_07730, D4F49_16455, D4F54_04680, D4F77_10190, D4F87_02985, D4F98_10640, D4G07_10045, D4G10_11575, D4G50_05510, D4X66_01450, D4X67_01465, D4X81_08350, D4Y13_13605, D4Y27_04430, D4Y38_00790, D4Y42_06150, D4Y56_03795, D4Y61_08865, D4Y96_04575, D4Z14_05470, D4Z16_07845, D4Z17_03400, D4Z29_11545, D4Z41_08675, D4Z50_07970, D4Z57_03410, D4Z58_04655, D4Z75_04825, D5764_07825, D5769_02495, D5773_00875, D5782_03660, D5810_01285, D5836_17200, D5848_11770, D5856_03150, D5861_06225, D5881_04345, D5889_07675, D5895_01345, D5905_18015, D5915_03200, D5970_05735, D5972_06010, D5A61_03660, D5A65_05805, D5A76_04875, D5A82_03865, D5A94_12405, D5B23_01450, D5B60_01340, D5B66_05165, D5B94_03135, D5C00_05295, D5C12_08175, D5C21_07220, D5C32_06325, D5C34_10465, D5C40_07860, D5C41_08390, D5C46_12375, D5C64_03810, D5C69_08580, D5O45_04325, D5O48_05730, D5O74_04490, D5O78_09935, D5O80_07835, D5O86_11185, D5P09_09970, D5P22_01470, D5P31_04355, D5P36_04640, D5P42_03925, D5P52_00720, D5P54_05700, D5P59_05915, D5P71_08860, D5P75_03560, D5P94_16990, D5Q05_03935, D5Q09_06085, D5Q39_11635, D5Q43_00445, D5W53_06130, D5W81_01065, D5W91_06810, D5X16_03865, D5X20_07410, D5X39_00990, D5X56_02780, D5X61_00680, D5X72_03440, D5X88_07550, D5X91_04060, D5Y01_00825, D5Y06_08270, D5Y08_23240, D5Y41_01425, D6331_01310, D6379_03305, D6389_04215, D6413_06485, D6429_04170, D6448_15635, D6453_04205, D6455_06220, D6467_08685, D6J43_21925, D6J46_01025, D6J55_02800, D6J67_02330, D6J74_05385, D6J84_08940, D6K21_03765, D6K24_01080, D6K59_04410, D6L05_07630, D6P24_03240, D6P41_01305, D6P57_11975, D6P63_03960, D6Q07_11960, D6Q21_01005, D6Q25_01020, D6Q70_23040, D6Q87_10085, D6Q96_09190, D6Q97_04185, D6R14_09640, D6R58_05020, D6R62_09555, D6R98_02810, D6S02_04705, D6S13_01360, D6S22_05290, D6S36_05340, D6S45_01045, D6S47_07470, D6S50_05845, D6S52_07325, D6S74_01450, D6S83_06345, D6S89_03890, D6T18_03025, D6T31_08100, D6T37_07430, D7M89_05935, D7M98_11725, D7N03_02455, D7N07_08155, D7N22_06745, D7N26_22710, D7N30_05075, D7N47_05845, D7N51_15930, D7N52_04690, D7N80_02885, D7N86_17235, D7O13_06330, D7O17_07925, D7O23_11190, D7O25_01030, D7O28_08970, D7O59_06225, D7O77_04615, D8Q64_01020, D8Q73_05540, D8R14_01045, D8R18_08390, D8R62_10510, D8R78_07850, D8S31_02190, D8S34_08660, D8S35_08175, D8S37_06530, D8S38_08610, D8S62_04305, D8S66_05780, D9O60_02875, D9O71_05360, D9O80_03040, D9O86_13290, D9O95_03895, D9P04_07965, D9P12_05975, D9P13_14425, D9P19_07675, D9P28_09530, D9P30_03370, D9P43_10815, D9P51_02955, D9P55_07730, D9P58_06870, D9P68_09885, D9P88_00985, D9P92_02905, D9P97_06685, D9Q07_09260, D9Q11_05725, D9Q23_03710, D9Q29_06205, D9Q37_07930, D9Q49_08285, D9Q50_10325, DAR35_01005, DAW17_06730, DAX91_07060, DFQ85_05430, DFR01_09160, DK013_01025, DK030_06300, DK052_23015, DK083_04615, DK093_01040, DK094_01025, DK101_08130, DK113_08305, DK117_07485, DK127_09260, DK678_00070, DK681_09420, DK687_04055, DK711_01855, DK719_05700, DK739_06740, DK741_10685, DK756_03460, DK767_12285, DKI70_07030, DKJ03_09570, DKJ07_05835, DKS84_08230, DKT00_02710, DKT04_15920, DKT21_12805, DKT27_10265, DKT52_07220, DKU65_14915, DKV05_06000, DKV20_06200, DKV25_04445, DL088_11075, DL104_17055, DL110_05250, DL111_06640, DL115_01355, DL130_03140, DL146_02805, DLB02_07650, DLB18_08505, DLB23_01000, DLB36_10070, DLB43_07510, DLB46_07880, DLB62_03890, DLB73_00105, DLC24_04935, DLE90_18415, DLF22_04805, DLF29_05020, DLF30_08180, DLF33_10805, DLF35_01005, DLF36_02845, DLM06_04560, DLM09_00995, DLM10_09965, DLM30_04930, DLQ73_12300, DLQ79_01000, DLQ86_15735, DLQ92_03590, DLR05_06915, DLR06_01490, DLR25_05290, DLR57_10745, DM001_07440, DM006_09100, DM016_05255, DM323_06700, DM347_08405, DM358_08140, DM367_10780, DM371_02430, DM700_18145, DM703_02460, DM705_08310, DM743_09605, DMA85_03170, DMA89_15850, DMB03_08830, DMB04_15185, DML85_00605, DML97_09050, DMM16_01960, DMM17_11150, DMM22_02845, DMM24_11405, DMM43_07890, DMO61_03735, DMO65_01050, DMO79_09060, DMU74_10415, DMU80_09050, DMU82_12600, DMU94_09600, DMV06_03650, DMV13_03350, DMV20_20060, DMV32_00825, DMV34_02950, DMZ76_01020, DN020_06485, DN087_03575, DN092_04545, DN123_07690, DN125_01440, DN130_05830, DN132_04380, DN181_04005, DN182_06745, DN220_06100, DN241_03340, DN257_05145, DN259_05090, DN271_19320, DN282_15665, DN317_01415, DN319_10795, DN322_05135, DN358_10690, DN362_06885, DNI00_08045, DNI25_05135, DNI46_06020, DNI61_09285, DNI77_05260, DNI82_12125, DNI85_03675, DNJ05_06165, DNJ30_02305, DNL79_03545, DNM18_05050, DNM22_08605, DNM39_12310, DNM44_03995, DNM47_08685, DNM48_03895, DNM80_06910, DNN00_05420, DNU17_09140, DNU22_04660, DNU30_01285, DNU40_06770, DNU46_14660, DNU67_08335, DNU68_11160, DNU78_13165, DNU82_03725, DNU84_13710, DNU93_05320, DNV11_03960, DNV18_07230, DNV24_12015, DNV55_05965, DNV93_02810, DNW15_00865, DNY10_04300, DNY38_00420, DNY86_09760, DNZ14_01330, DNZ24_04550, DNZ41_01315, DNZ69_08840, DNZ74_07090, DNZ75_08705, DNZ81_03425, DNZ91_05745, DO557_05890, DO565_08085, DO591_06685, DO596_04935, DO617_04810, DO619_02490, DO630_04000, DO638_05650, DO649_13400, DO654_06280, DO661_06270, DO663_04365, DO672_01035, DO679_08555, DO681_04055, DO690_00395, DO696_07715, DO712_22550, DO727_09960, DO731_01530, DO771_06730, DO807_07800, DO827_06795, DO973_04300, DO981_01690, DO994_01335, DOA98_08020, DOB00_09740, DOB23_01440, DOB53_16610, DOB60_05740, DOB76_01635, DOC12_00300, DOC15_05765, DOC25_09930, DOC31_08415, DOC34_09315, DOC40_14960, DOC63_04280, DOG94_02935, DOH04_12155, DOH12_22545, DOH33_03235, DOH41_03550, DOH63_02830, DOH69_10710, DOH74_13625, DOH75_06000, DOH81_09930, DOI30_01020, DOI60_10480, DOI66_05845, DOJ02_05395, DOQ45_03150, DOQ73_06415, DOQ79_01065, DOR42_03375, DOR43_04775, DOR46_16150, DOR51_23780, DOR52_02535, DOR63_07395, DOR64_10515, DOR75_06845, DOV68_03030, DOV71_04785, DOV77_15350, DOV89_01625, DOW27_10165, DOW52_22585, DOW74_04075, DOX21_02055, DOX30_03420, DP688_13895, DP693_00825, DP698_06830, DP703_05145, DP712_02970, DP713_03380, DP716_06840, DP727_22840, DP728_16275, DP736_07365, DP748_02490, DP766_03155, DP778_03250, DP792_05710, DP800_16220, DP812_13225, DP837_03195, DP859_02865, DP873_09820, DP884_06665, DPA01_04065, DPA17_01430, DPA21_08565, DPA36_04755, DPA81_08695, DPA91_07750, DPB01_05745, DPB58_05530, DPB61_01065, DPB73_06705, DPB80_01010, DPC17_04415, DPC26_12120, DPC30_09225, DPC31_21295, DPC91_07340, DPD28_03410, DPD35_09350, DPD61_05565, DPD71_12990, DPD96_04945, DPE17_03960, DPE32_06850, DPE40_05345, DPE61_01100, DPE66_10470, DPE72_00995, DPE76_10585, DPE91_07335, DPE98_01700, DPF18_01415, DPF55_03280, DPF57_13210, DPF65_05290, DPF69_04225, DPJ06_06690, DPJ07_04770, DPJ18_21730, DPJ21_23060, DPJ65_05985, DPJ69_12010, DPJ81_01030, DPK19_02945, DPK49_18945, DPL03_00940, DPP06_22450, DPP34_02995, DPP49_05140, DPP57_09185, DPP70_04625, DPR96_17095, DPS51_01190, DPS62_02400, DPS71_04035, DPS75_05935, DPS77_06815, DPS96_17080, DPT22_03505, DPT41_07335, DPT74_05620, DPT84_17600, DPT95_03965, DPU11_05080, DPU13_04230, DPY66_15080, DPY81_09375, DPZ06_03150, DPZ23_03295, DPZ65_01920, DPZ67_03455, DPZ88_09965, DPZ95_04280, DQ003_10345, DQ017_21125, DQ037_03770, DQ055_06250, DQ072_13750, DQ076_16795, DQ078_07335, DQ082_06505, DQ790_02530, DQ802_06935, DQ816_09465, DQ836_21070, DQ837_05050, DQ839_09850, DQ845_10875, DQ858_00055, DQ871_15205, DQ880_07325, DQ895_06115, DQ901_09380, DQ919_02340, DQ933_07675, DQ978_14715, DQ985_10135, DQC12_07750, DQC17_05135, DQC26_03870, DQC75_03590, DQC77_06245, DQC81_16925, DQC82_09655, DQC88_01435, DQD09_24240, DQD11_01060, DQD13_03865, DQD28_13095, DQD46_02455, DQD59_03945, DQE08_00395, DQE20_03160, DQE31_05590, DQE33_13015, DQE40_13690, DQE42_08765, DQJ22_01340, DQJ50_08905, DQJ58_03830, DQJ63_05540, DQJ66_02440, DQJ71_08430, DQJ77_01030, DQJ83_05310, DQJ90_08450, DQK19_10845, DQK43_01440, DQK45_13610, DQK53_05425, DQK57_20355, DQK72_09740, DQQ27_15440, DQR39_03355, DQR40_15910, DQR74_07115, DQR85_07280, DQR90_05875, DQS34_02500, DQS55_04230, DQS56_07240, DQS62_04550, DQS71_07420, DQS74_06295, DQS89_04330, DQS98_04035, DQT21_23885, DQT31_01485, DQT40_01975, DQT47_08080, DQT59_08365, DQT67_04950, DQT85_05480, DQY62_10275, DQY86_01365, DQY91_07170, DQZ13_08220, DQZ68_04575, DQZ89_03655, DRA21_00825, DRA23_02570, DRA25_06475, DRA26_15005, DRA30_07960, DRA37_10395, DRD38_06525, DRD40_04685, DRD55_03550, DRD57_02825, DRD79_08175, DRE09_03295, DRE12_04035, DRE16_07695, DRE32_01375, DRE41_16345, DRE46_06240, DRE58_03660, DRE69_05135, DRE85_05775, DRE92_01430, DRF10_01070, DRF26_07295, DRF38_06650, DRK15_08305, DRK36_10935, DRK49_10210, DRK71_14235, DRK91_06225, DRL06_03150, DRL40_04985, DRL53_01550, DRL62_15150, DRL63_08155, DRL65_05050, DRL68_04235, DRL89_01035, DRL96_01185, DRM19_02850, DRM39_10070, DRM41_05515, DRM43_02825, DRM75_09425, DRT10_03415, DRT15_04045, DRT45_11245, DRT49_10675, DRT56_11825, DRT73_05670, DRT76_09345, DRT77_01030, DRU07_04960, DRU14_01025, DRU25_12475, DRU99_06930, DRV08_07615, DRV17_01045, DRV44_09955, DRV53_06735, DRV60_09675, DRV62_04555, DRW94_06120, DRX19_09970, DRX40_11405, DRX43_00440, DRY30_09095, DRY35_08945, DRY53_03870, DRY54_04540, DRY73_06120, DRZ18_11450, DS161_03605, DS168_01430, DS172_06735, DS181_05705, DS188_01805, DS209_03915, DS212_06790, DS228_01025, DS242_04605, DS251_06835, DS266_01340, DS269_03610, DS300_04175, DS306_05655, DS313_01340, DS324_09265, DS336_04600, DS340_09465, DS346_12760, DS368_20110, DS372_11155, DS390_10475, DS438_03885, DS468_03185, DS470_17635, DS485_05030, DS497_03915, DS570_05955, DS572_04860, DS605_10670, DS615_10365, DS651_15910, DS657_02820, DS688_11950, DS689_13215, DS708_08160, DSA82_08035, DSA90_01010, DSA91_03875, DSA99_10695, DSB20_06505, DSB26_11025, DSB29_01055, DSB36_03930, DSB44_04370, DSE94_03270, DSF03_05385, DSF06_15125, DSF13_00100, DSF32_06125, DSF57_04690, DSF62_14785, DSF92_08520, DSF93_06370, DSG01_03865, DSG07_07830, DSG23_03190, DSG69_22660, DSG96_09425, DSH12_03820, DSM73_23085, 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F3E81_09345, F3P41_00915, F3P70_08045, F3P71_01925, F3P74_06490, F3P75_12190, F3P83_04915, F3Q00_03840, F3Q03_05030, F3Q14_02890, F3Q15_02920, F3Q17_04825, F3Q19_00915, F3Q28_06760, F3Q63_04915, F3R19_08235, F3X32_06195, F3X39_18195, F3X53_04150, F3X54_06825, F3X74_09220, F6430_00280, F7247_03195, F7801_03470, F9P04_11415, F9P26_00380, F9Q85_11085, F9W02_05115, F9X76_16290, F9X98_08000, FE718_01360, FE738_01015, FE759_06665, FKN96_12870, FL809_10360, FMV10_12745, FP220_08405, FPF85_03010, FQS21_06835, FQS30_06725, FQS47_00615, FQS59_09325, FR245_13140, FR247_08285, FRM70_05500, FRM79_14805, FRN01_11890, FRN02_05030, FRN09_04175, FRN22_13530, FY061_02260, FZ484_04550, FZ491_03810, FZ502_10420, FZ514_03745, FZ515_13330, FZ516_05975, FZ523_01470, FZ527_01765, FZ528_04325, FZT79_05320, GAW88_10720, GCH48_00965, GCX84_03060, GCY28_22200, GCY29_11985, GDH50_05360, GDH65_00960, GN92_19330, GX90_25025, JF03_19690, JR37_17475, KO25_20305, LB54_08610, LC09_03640, LC85_05730, LH24_08730, LH82_14165, LP13_20040, MB64_17020, NB08_11105, NCTC8272_02769, NU30_22415, PK04_18245, PK95_13970, PV65_06390, QC88_19815, QD42_08685, QD82_16765, QE89_08685, QE93_10380, QF21_24215, R118_18445, R126_08305, R133_19845, RU16_07410, RU46_12145, RY46_16045, RY52_16035, S021_09745, S313_01135, S315_02435, S598_12875, S639_21410, S709_07710, S764_16455, S943_05250, TE27_11430, TL61_17385, VT29_18010, WB97_02550, XJ41_07365, XK54_01525, XK85_16820, XS28_05335, XT48_02390, XZ16_15885, Y847_07225, YA74_10200, YH39_16290, YI72_11450, YJ27_07680, YL04_09955, YL55_19460, YR14_06075, YU10_04620, Z155_07555, Z599_02180, ZA28_00110, ZA51_00590, ZA74_09315, ZD91_01440, ZF07_21535, ZG82_06210, ZO67_01860
Production host: Escherichia coli (E. coli) / References: UniProt: A0A3U3J7F4, UniProt: Q8ZRA7*PLUS

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentAggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction

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Sample preparation

ComponentName: AcrBAcriflavine resistance protein family / Type: COMPLEX / Entity ID: all / Source: RECOMBINANT
Molecular weightExperimental value: NO
Source (natural)Organism: Salmonella enterica subsp. enterica serovar Typhi (bacteria)
Source (recombinant)Organism: Escherichia coli (E. coli)
Buffer solutionpH: 7.5
SpecimenConc.: 1 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
VitrificationCryogen name: ETHANE

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyModel: FEI TITAN KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM
Electron lensMode: BRIGHT FIELDBright-field microscopy
Image recordingElectron dose: 60 e/Å2 / Film or detector model: FEI FALCON III (4k x 4k)

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Processing

SoftwareName: PHENIX / Version: 1.12_2829: / Classification: refinement
CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
SymmetryPoint symmetry: C3 (3 fold cyclic)
3D reconstructionResolution: 4.6 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 316000 / Symmetry type: POINT
Refine LS restraints
Refine-IDTypeDev idealNumber
ELECTRON MICROSCOPYf_bond_d0.00645213
ELECTRON MICROSCOPYf_angle_d0.72981251
ELECTRON MICROSCOPYf_dihedral_angle_d7.06218192
ELECTRON MICROSCOPYf_chiral_restr0.0493788
ELECTRON MICROSCOPYf_plane_restr0.0056989

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