2FZM

Structure of the E. coli PutA proline dehydrogenase domain reduced by dithionite and complexed with SO2

2FZM の概要

• エントリーDOI: 10.2210/pdb2fzm/pdb
• 分子名称: Bifunctional protein putA, Proline dehydrogenase (EC 1.5.99.8) (Proline oxidase), FLAVIN-ADENINE DINUCLEOTIDE, SULFUR DIOXIDE, ... (4 entities in total)
• 機能のキーワード: proline utilization a, proline dehydrogenase, puta, flavoenzyme, proline catabolism, dithionite-reduced, oxidoreductase
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 67517.68

構造登録者

Tanner, J.J. (登録日: 2006-02-09, 公開日: 2007-04-03, 最終更新日: 2023-08-30)

主引用文献

Zhang, W.,Zhang, M.,Zhu, W.,Zhou, Y.,Wanduragala, S.,Rewinkel, D.,Tanner, J.J.,Becker, D.F.
Redox-induced changes in flavin structure and roles of flavin N(5) and the ribityl 2'-OH group in regulating PutA--membrane binding.
Biochemistry, 46:483-491, 2007

PubMed Abstract: PutA is a novel flavoprotein in Escherichia coli that switches from a transcriptional repressor to a membrane-bound proline catabolic enzyme. Previous crystallographic studies of the PutA proline dehydrogenase (PRODH) domain under oxidizing conditions revealed that FAD N(5) and the ribityl 2'-OH group form hydrogen bonds with Arg431 and Arg556, respectively. Here we identify molecular interactions in the PutA PRODH active site that underlie redox-dependent functional switching of PutA. We report that reduction of the PRODH domain induces major structural changes in the FAD cofactor, including a 22 degrees bend of the isoalloxazine ring along the N(5)-N(10) axis, crankshaft rotation of the upper part of the ribityl chain, and formation of a new hydrogen bond network involving the ribityl 2'-OH group, FAD N(1), and Gly435. The roles of the FAD 2'-OH group and the FAD N(5)-Arg431 hydrogen bond pair in regulating redox-dependent PutA-membrane associations were tested using FAD analogues and site-directed mutagenesis. Kinetic membrane binding measurements and cell-based reporter gene assays of modified PutA proteins show that disrupting the FAD N(5)-Arg431 interaction impairs the reductive activation of PutA-membrane binding. We also show that the FAD 2'-OH group acts as a redox-sensitive toggle switch that controls PutA-membrane binding. These results illustrate a new versatility of the ribityl chain in flavoprotein mechanisms.

PubMed: 17209558
DOI: 10.1021/bi061935g

実験手法

X-RAY DIFFRACTION (2.3 Å)