2021-06-30RELPDBjPDBjPDBj2020-08-112021-04-142021-04-142021-06-30Japan Society for the Promotion of Science (JSPS)19H04645JapanJapan Agency for Medical Research and Development (AMED)JP20am0101071JapanCryo-EM analysis of the nonribosomal peptide synthetase, FmoA3Sone KHarada AKawai SUrano NAdachi NMoriya TKawasaki MKatsuyama YSenda TOhnishi YKatsuyama YSone KHarada AKawai SUrano NAdachi NMoriya TKawasaki MShin-Ya KSenda TOhnishi YStructural and Functional Analyses of the Tridomain-Nonribosomal Peptide Synthetase FmoA3 for 4-Methyloxazoline Ring Formation.Angew.Chem.Int.Ed.Engl.GE601455414562202133783097doi:10.1002/anie.2021027601521-37730179ACIEAYEMD-30440associated EM volumeStructure of nonribosomal peptide synthetase, FmoA3Structure of nonribosomal peptide synthetase, FmoA301Streptomyces sp. Sp080513GE-230.12Escherichia coli BL21(DE3)BL21pColdIFmoA3Streptomyces sp. Sp080513GE-23Escherichia coli BL21(DE3)LEVOMTISVPGHGTLGAPDGAAAPGGEAAELPPLVPEPGDAGQPFPLTPTQQALWVGRADAVDLGDIGCYGYFE
WERPELDLARYRRAWERLVAHHPGLRTVVRPDGTQHVLERPGPVPITVEDLRQDPDAVRRLEESRAERGH
RALDPGTWPMFDLRVVLLSGRVRVQLGIDLQLMDASSLFLNLFSDLVTLYDDPDAALASQKLAFRDFARW
LEEDVRGGARWRADWAYWQERLDGLPPAPDLPAARYGAQGPGKFERCMVRCPAEEFALLRERALAHGLTE
TELLVGAFAEVLRGWSSDPAFTLNVPVFQRFDVPGIEDVIGDYTNPILLEARPEGRTVAERIVALAARLR
ADTRHASVNGVEVLRELARRRGLAAAAMPVVVTSLLGLPSAARSITEFGTEVHSITQTPQVSLDFQIRPE
DGELRLVWDHRSGAFAPGVVEGAFEAFLDLVGRMLADEPGHGVWEAPFADMRSRRDRAVWNETNDTAEPV
PAVLLQERFFAQARRTPDAEAVVASGLRLTYDELARHAYRIGNTLRERGVRPGDLVGVVMEKGWEQYAAV
YGILAAGGAYLPIDAASPRGRVARLLESAGAGIVLTQSRLRDELDLPAGTTVLRADTDFETASTAPLTPV
QGPDDPAYVIYTSGSTGEPKGVVVAHRGVANLVRDVRRRFAVTPADRLLALSGLHFDASVYDVFGPLACG
ATVVVPPPFRRAEPDVWAELVRDERVTFWNSVPVLLELLVGEAESRDDRPLATLRLAVVSGDWIPLDLPG
RARAQAPGLRVVGSGGPTETICWSLFHPIDAVDPQWTSIPYGKPIANQRYYIVDRDLRPRPTWARGEMAV
ASPLGLALGYLNDPERTAAKFVTLPGTGERAYLTGDFGRLLPDGGIEILGREDFQVKVAGQRIELGEIEA
LLHRADGVRAAVVTAPRSSADVVRLQAFVVPETGARLSADALREHLSAELPAAMVPAAIRLLPELPLTAN
GKVDRLALARLAAAPEEAPEPEARTDYAPRTDVGLLAELVAACVAELLGLDEVPTTGNFFRLGGDSLSGT
RLASRLQDLLGAPVPIRTVFGNPVLGDLASAIAGDPAAGPQAIRVARLLHTLEEPDEKPGEKPDAEPAGE
PDAGSRTsingleParticleparticle88.020.0C8H18N2O4SHEPES1.0C4H10O2S2DTTQuantifoil R1.2/1.3COPPER300CARBONHOLEYGLOW DISCHARGEAIRThe grid was washed by acetone prior to use.ETHANE100291FEI VITROBOT MARK IVBlotting time was 15 seconds (blot force 25). This sample was mono-disperse.FEI TALOS ARCTICAFLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN20050.02.71.02.5120000.0NITROGENFEI FALCON III (4k x 4k)COUNTING1188650.0750.01819697Gctf1.06An ab initio model was generated using RELION3's own implementation of Stochastic Gradient Descent (SGD) algorithm and low-pass filtered to 60 A for use as an initial model for 3D classification.C23.55FSC 0.143 CUT-OFFRELION376171MAXIMUM LIKELIHOODRELION3MAXIMUM LIKELIHOODRELION3emd_30440_half_map_1.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
480
480480
0
00480480480422.4422.4422.490.090.090.0XYZ-0.03772090.087901281.6453247e-050.00276831980.880.880.88emd_30440_half_map_2.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)