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

Basic information

Entry

Database: SASBDB / ID: SASDDY6

Sample

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

Function / homology

Function:

photosynthetic acclimation / photosynthetic electron transport chain / chloroplast / 2 iron, 2 sulfur cluster binding / metalloendopeptidase activity / electron transfer activity / proteolysis / metal ion binding

Domain/homology:

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

Component:

Ferredoxin-2, chloroplastic / Zinc protease

• Biological species: Pectobacterium atrosepticum SCRI1043 (bacteria); Arabidopsis thaliana (thale cress)
• Citation: Journal: PLoS Biol / Year: 2018; Title: FusC, a member of the M16 protease family acquired by bacteria for iron piracy against plants.; Authors: 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 / ; Abstract: 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.

Contact author

• Grinter Grinter (Monash University, Melbourne, Australia)

Models

Sample

• Sample: Name: The ferredoxin protease, FusC, E83A mutant + 20 µM Arabidopsis ferredoxin; Specimen concentration: 3 mg/ml / Entity id: 1084 / 1085
• Buffer: Name: 20 mM Tris, 150 mM NaCl / pH: 7.8

Entity #1084

Name: FusC E83A / Type: protein / Description: Ferredoxin protease E83A mutant / Formula weight: 101.195 / Num. of mol.: 1 / Source: Pectobacterium atrosepticum SCRI1043 / References: UniProt: Q6D8U3
Sequence:

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

Entity #1085

Name: Ara_Fer2 / Type: protein / Description: Arabidopsis ferredoxin 2 / Formula weight: 11.33 / Num. of mol.: 1 / Source: Arabidopsis thaliana / References: UniProt: P16972
Sequence:

ATYKVKFITP EGELEVECDD DVYVLDAAEE AGIDLPYSCR AGSCSSCAGK VVSGSVDQSD QSFLDDEQIG EGFVLTCAAY PTSDVTIETH KEEAIMLEHH HHHH

Experimental information

• Beam: Instrument name: Australian Synchrotron SAXS/WAXS / City: Melbourne / 国: Australia / Shape: Point / Type of source: X-ray synchrotronSynchrotron / Wavelength: 0.103 Å / Dist. spec. to detc.: 1.28 mm
• Detector: Name: Pilatus 1M / Pixsize x: 172 mm

Scan

Title: The ferredoxin protease, FusC, E83A mutant + 20 µM Arabidopsis ferredoxin
Measurement date: Oct 26, 2017 / Storage temperature: 20 °C / Cell temperature: 20 °C / Unit: 1/A /

	Min	Max
Q	0.0114	0.3038

Distance distribution function P(R)

Sofotware P(R): GNOM 5.0 / Number of points: 273 /

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

Result

Type of curve: single_conc
Comments: The FusC E83A mutant (30 µM) in the presence of 20 µM Arabidopsis ferredoxin. The background subtraction included 20 µM Arabidopsis ferredoxin in buffer.

	Experimental	Porod
MW	100 kDa	91 kDa
Volume	-	154.9 nm3

	P(R)	Guinier	Guinier error
Forward scattering, I0	0.2699	0.28	0.00022
Radius of gyration, Rg	3.76 nm	3.68 nm	0.025

	Min	Max
D	-	13.62
Guinier point	9	32