SASDDY6: The ferredoxin protease, FusC, E83A mutant + 20 µM Arabidopsis fe...

基本情報

登録情報

データベース: SASBDB / ID: SASDDY6

試料

The ferredoxin protease, FusC, E83A mutant + 20 µM Arabidopsis ferredoxin

• Ferredoxin protease E83A mutant (protein), FusC E83A, Pectobacterium atrosepticum SCRI1043
• Arabidopsis ferredoxin 2 (protein), Ara_Fer2, Arabidopsis thaliana

機能・相同性

分子機能:

photosynthetic acclimation / photosynthetic electron transport chain / 葉緑体 / 2 iron, 2 sulfur cluster binding / metalloendopeptidase activity / electron transfer activity / タンパク質分解 / metal ion binding

ドメイン・相同性:

Ferredoxin [2Fe-2S], plant / 2Fe-2S ferredoxin, iron-sulphur binding site / 2Fe-2S ferredoxin-type iron-sulfur binding region signature. / Peptidase M16, zinc-binding site / Insulinase family, zinc-binding region signature. / 2Fe-2S iron-sulfur cluster binding domain / Peptidase M16, C-terminal / Peptidase M16 inactive domain / Peptidase M16, N-terminal / Insulinase (Peptidase family M16) / Metalloenzyme, LuxS/M16 peptidase-like / Beta-grasp domain superfamily / 2Fe-2S ferredoxin-type iron-sulfur binding domain profile. / 2Fe-2S ferredoxin-type iron-sulfur binding domain / 2Fe-2S ferredoxin-like superfamily

構成要素:

Ferredoxin-2, chloroplastic / Zinc protease

• 生物種: Pectobacterium atrosepticum SCRI1043 (バクテリア); Arabidopsis thaliana (シロイヌナズナ)
• 引用: ジャーナル: PLoS Biol / 年: 2018; タイトル: FusC, a member of the M16 protease family acquired by bacteria for iron piracy against plants.; 著者: Rhys Grinter / Iain D Hay / Jiangning Song / Jiawei Wang / Don Teng / Vijay Dhanesakaran / Jonathan J Wilksch / Mark R Davies / Dene Littler / Simone A Beckham / Ian R Henderson / Richard A Strugnell / Gordon Dougan / Trevor Lithgow / ; 要旨: Iron is essential for life. Accessing iron from the environment can be a limiting factor that determines success in a given environmental niche. For bacteria, access of chelated iron from the environment is often mediated by TonB-dependent transporters (TBDTs), which are β-barrel proteins that form sophisticated channels in the outer membrane. Reports of iron-bearing proteins being used as a source of iron indicate specific protein import reactions across the bacterial outer membrane. The molecular mechanism by which a folded protein can be imported in this way had remained mysterious, as did the evolutionary process that could lead to such a protein import pathway. How does the bacterium evolve the specificity factors that would be required to select and import a protein encoded on another organism's genome? We describe here a model whereby the plant iron-bearing protein ferredoxin can be imported across the outer membrane of the plant pathogen Pectobacterium by means of a Brownian ratchet mechanism, thereby liberating iron into the bacterium to enable its growth in plant tissues. This import pathway is facilitated by FusC, a member of the same protein family as the mitochondrial processing peptidase (MPP). The Brownian ratchet depends on binding sites discovered in crystal structures of FusC that engage a linear segment of the plant protein ferredoxin. Sequence relationships suggest that the bacterial gene encoding FusC has previously unappreciated homologues in plants and that the protein import mechanism employed by the bacterium is an evolutionary echo of the protein import pathway in plant mitochondria and plastids.

登録者

• Grinter Grinter (Monash University, Melbourne, Australia)

モデル

試料

• 試料: 名称: The ferredoxin protease, FusC, E83A mutant + 20 µM Arabidopsis ferredoxin; 試料濃度: 3 mg/ml / Entity id: 1084 / 1085
• バッファ: 名称: 20 mM Tris, 150 mM NaCl / pH: 7.8

要素 #1084

名称: FusC E83A / タイプ: protein / 記述: Ferredoxin protease E83A mutant / 分子量: 101.195 / 分子数: 1 / 由来: Pectobacterium atrosepticum SCRI1043 / 参照: UniProt: Q6D8U3
配列:

EEIKSPLPVF KEGTLANGFR YTLVQLEGPK TRVDIRLIVD VGSIDEKDNE SGVAHMVAHM VFRASDAFPQ GVSTELHKQG WGRGQSYNAV TNYERTMYMM SPPKGNLDLG ATLQALSQMT GHAKLLQSDL DDERKIILEE WRGKLGVAER MNQQRVQAIR HDSRYPSRPV IGTEESINDT PASVLQDFYQ RWYHPSNMRL MIIGDITPAD AEREIQRYFA ALPNVAVPTR DYYEPLLKPQ LKVARLQDSQ SGSSQVSFVY RFNDKDAFGQ SEYRHRLLTQ ITMSAVTRQV RRQKAELPQD ASSLVVRKSD IGKTTAALGF FANVMPGGHD AAISAVLKEI ERFKRYPLNE QDITEITSDI REVAQRMSVT PETREFADWV QQLTIVWQQD RPYVGSQQRG KDALEALDTI KGEDVNRHWQ RWLASPDTLA QFSVPGATPF TLPKPDAISK LQKQWALATL APLRLEEKKI IPELPSVTQS GKRTAVKTFA AQKVEQWQLS NGDRVVWLRA PEAGKKVYLT ATSQAGFMAT AMNPWQAQLA SQLVNQSGPA TWSGESLSNW KKEKTLSLSI DQEADQLTLS GTAPTEQLAS LFGLYRELNV APGIDPDVMK ESMMSLARQK ANDDQSVGGK RASEMTKLRF GEPAWQQPEI AELKKISAPA LLSQWHKAAS APVTYYLIAD MPATQLLPQV ERYLATIPRQ PASEVKQHLA LSGKREATSA INVEPRADIL TWSFTPHAWT PQAAVQVSIA RNIASKYLKT SLRDDALGIY RMRVDSELED KKQRIETEVS FTSAPERAQE LWTLAEQAFS ELPTKITQQD VDEQKAQFIR AEKGRQGDLT TIQRRLILSY RHYNDPRYLS NASKLADSIT LESVRAMSAK LYNPDNRVLY ITLPQEVKE

要素 #1085

名称: Ara_Fer2 / タイプ: protein / 記述: Arabidopsis ferredoxin 2 / 分子量: 11.33 / 分子数: 1 / 由来: Arabidopsis thaliana / 参照: UniProt: P16972
配列:

ATYKVKFITP EGELEVECDD DVYVLDAAEE AGIDLPYSCR AGSCSSCAGK VVSGSVDQSD QSFLDDEQIG EGFVLTCAAY PTSDVTIETH KEEAIMLEHH HHHH

実験情報

• ビーム: 設備名称: Australian Synchrotron SAXS/WAXS / 地域: Melbourne / 国: Australia / 形状: Point / 線源: X-ray synchrotronシンクロトロン / 波長: 0.103 Å / スペクトロメータ・検出器間距離: 1.28 mm
• 検出器: 名称: Pilatus 1M / Pixsize x: 172 mm

スキャン

タイトル: The ferredoxin protease, FusC, E83A mutant + 20 µM Arabidopsis ferredoxin
測定日: 2017年10月26日 / 保管温度: 20 °C / セル温度: 20 °C / 単位: 1/A /

	Min	Max
Q	0.0114	0.3038

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 273 /

	Min	Max
Q	0.0172374	0.216648
P(R) point	1	273
R	0	136.2

結果

カーブのタイプ: single_conc
コメント: The FusC E83A mutant (30 µM) in the presence of 20 µM Arabidopsis ferredoxin. The background subtraction included 20 µM Arabidopsis ferredoxin in buffer.

	実験値	Porod
分子量	100 kDa	91 kDa
体積	-	154.9 nm3

	P(R)	Guinier	Guinier error
前方散乱 I0	0.2699	0.28	0.00022
慣性半径, Rg	3.76 nm	3.68 nm	0.025

	Min	Max
D	-	13.62
Guinier point	9	32