4RZT

Lac repressor engineered to bind sucralose, sucralose-bound tetramer

4RZT の概要

• エントリーDOI: 10.2210/pdb4rzt/pdb
• 関連するPDBエントリー: 4RZS
• 関連するBIRD辞書のPRD_ID: PRD_900090
• 分子名称: Lac repressor, 4-chloro-4-deoxy-alpha-D-galactopyranose-(1-2)-1,6-dichloro-1,6-dideoxy-beta-D-fructofuranose (3 entities in total)
• 機能のキーワード: sucralose, laci, lac repressor, lactose operon repressor, protein design, allostery, escherichia coli, transcription
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 165124.49

構造登録者

Arbing, M.A.,Cascio, D.,Sawaya, M.R.,Kosuri, S.,Church, G.M. (登録日: 2014-12-24, 公開日: 2015-12-23, 最終更新日: 2024-02-28)

主引用文献

Taylor, N.D.,Garruss, A.S.,Moretti, R.,Chan, S.,Arbing, M.A.,Cascio, D.,Rogers, J.K.,Isaacs, F.J.,Kosuri, S.,Baker, D.,Fields, S.,Church, G.M.,Raman, S.
Engineering an allosteric transcription factor to respond to new ligands.
Nat.Methods, 13:177-183, 2016

PubMed Abstract: Genetic regulatory proteins inducible by small molecules are useful synthetic biology tools as sensors and switches. Bacterial allosteric transcription factors (aTFs) are a major class of regulatory proteins, but few aTFs have been redesigned to respond to new effectors beyond natural aTF-inducer pairs. Altering inducer specificity in these proteins is difficult because substitutions that affect inducer binding may also disrupt allostery. We engineered an aTF, the Escherichia coli lac repressor, LacI, to respond to one of four new inducer molecules: fucose, gentiobiose, lactitol and sucralose. Using computational protein design, single-residue saturation mutagenesis or random mutagenesis, along with multiplex assembly, we identified new variants comparable in specificity and induction to wild-type LacI with its inducer, isopropyl β-D-1-thiogalactopyranoside (IPTG). The ability to create designer aTFs will enable applications including dynamic control of cell metabolism, cell biology and synthetic gene circuits.

PubMed: 26689263
DOI: 10.1038/nmeth.3696

実験手法

X-RAY DIFFRACTION (3.1 Å)