Centre National de la Recherche Scientifique (CNRS)
Institute for Integrative Biology of the Cell (I2BC)
European Institute of Chemistry and Biology (IECB)
引用
ジャーナル: Sci Adv / 年: 2021 タイトル: Architecture and regulation of an enterobacterial cellulose secretion system. 著者: Wiem Abidi / Samira Zouhir / Meryem Caleechurn / Stéphane Roche / Petya Violinova Krasteva / 要旨: Many free-living and pathogenic enterobacteria secrete biofilm-promoting cellulose using a multicomponent, envelope-embedded Bcs secretion system under the control of intracellular second messenger c- ...Many free-living and pathogenic enterobacteria secrete biofilm-promoting cellulose using a multicomponent, envelope-embedded Bcs secretion system under the control of intracellular second messenger c-di-GMP. The molecular understanding of system assembly and cellulose secretion has been largely limited to the crystallographic studies of a distantly homologous BcsAB synthase tandem and a low-resolution reconstruction of an assembled macrocomplex that encompasses most of the inner membrane and cytosolic subunits and features an atypical layered architecture. Here, we present cryo-EM structures of the assembled Bcs macrocomplex, as well as multiple crystallographic snapshots of regulatory Bcs subcomplexes. The structural and functional data uncover the mechanism of asymmetric secretion system assembly and periplasmic crown polymerization and reveal unexpected subunit stoichiometry, multisite c-di-GMP recognition, and ATP-dependent regulation.
根拠: light scattering, SEC-MALS measurements confirmed the existence of a stable and monodisperse BcsR2Q2 heterotetramer in solution.
タイプ
名称
対称操作
数
identity operation
1_555
x,y,z
1
Buried area
12580 Å2
ΔGint
-90 kcal/mol
Surface area
20620 Å2
手法
PISA
単位格子
Length a, b, c (Å)
57.000, 78.990, 66.820
Angle α, β, γ (deg.)
90.000, 99.258, 90.000
Int Tables number
4
Space group name H-M
P1211
Space group name Hall
P2yb
Symmetry operation
#1: x,y,z #2: -x,y+1/2,-z
-
要素
#1: タンパク質
BacterialcellulosesecretionregulatorBcsQ / Cellulose biosynthesis protein BcsQ / Cellulose synthase operon protein YhjQ / Cellulose synthase / putative
分子量: 29245.209 Da / 分子数: 2 / 由来タイプ: 組換発現 詳細: Bacterial cellulose synthesis subunit Q. The coding region corresponding to the BcsRQ tandem was cloned into the pET21b expression vector with a C-terminal hexahistidine tag on BcsQ. The ...詳細: Bacterial cellulose synthesis subunit Q. The coding region corresponding to the BcsRQ tandem was cloned into the pET21b expression vector with a C-terminal hexahistidine tag on BcsQ. The protein complex was purified and crystallised in absence of nucleotide 由来: (組換発現) Escherichia coli (大腸菌) 遺伝子: bcsQ, yhjQ, A8C65_00290, ACU57_05335, BMT91_17060, BON75_10030, BvCmsHHP019_01723, BvCmsSINP011_05061, C2U48_15650, D3O91_11725, D9H68_14440, D9I18_15755, D9I97_13990, DAH34_22885, DAH37_ ...遺伝子: bcsQ, yhjQ, A8C65_00290, ACU57_05335, BMT91_17060, BON75_10030, BvCmsHHP019_01723, BvCmsSINP011_05061, C2U48_15650, D3O91_11725, D9H68_14440, D9I18_15755, D9I97_13990, DAH34_22885, DAH37_19450, E2127_16420, E2128_18010, E2129_18145, E2134_17810, EAI42_04085, EC1094V2_71, NCTC10429_00778, NCTC11022_03734, NCTC9058_01652 発現宿主: Escherichia coli BL21(DE3) (大腸菌) / 参照: UniProt: A0A0B1KWQ0, UniProt: P0DP92*PLUS
#2: タンパク質
BacterialcellulosesecretionregulatorBcsR / FIG004405: Putative cytoplasmic protein / Protein YhjR / Protein of uncharacterized function (DUF2629)