SASDCD2: Rab family protein CtRoc-COR GppNHp (Rab family protein)

Basic information

Entry

Database: SASBDB / ID: SASDCD2

Sample

Rab family protein CtRoc-COR GppNHp

• Rab family protein (protein), Chlorobium tepidum (strain ATCC 49652 / DSM 12025 / NBRC 103806 / TLS)

Function / homology

Function:

GTP binding / identical protein binding

Domain/homology:

C-terminal of Roc (COR) domain / C-terminal of Roc, COR, domain / Leucine rich repeat 4 / Leucine Rich repeats (2 copies) / Roc domain profile. / Roc domain / Leucine-rich repeat, typical subtype / Leucine-rich repeats, typical (most populated) subfamily / Leucine-rich repeat profile. / Leucine-rich repeat / Leucine-rich repeat domain superfamily / Small GTP-binding protein domain / Winged helix-like DNA-binding domain superfamily / P-loop containing nucleoside triphosphate hydrolase

Component:

Rab family protein

• Biological species: Chlorobium tepidum (strain ATCC 49652 / DSM 12025 / NBRC 103806 / TLS) (bacteria)
• Citation: Journal: Nat Commun / Year: 2017; Title: A homologue of the Parkinson's disease-associated protein LRRK2 undergoes a monomer-dimer transition during GTP turnover.; Authors: Egon Deyaert / Lina Wauters / Giambattista Guaitoli / Albert Konijnenberg / Margaux Leemans / Susanne Terheyden / Arsen Petrovic / Rodrigo Gallardo / Laura M Nederveen-Schippers / Panagiotis S Athanasopoulos / Henderikus Pots / Peter J M Van Haastert / Frank Sobott / Christian Johannes Gloeckner / Rouslan Efremov / Arjan Kortholt / Wim Versées / ; Abstract: Mutations in LRRK2 are a common cause of genetic Parkinson's disease (PD). LRRK2 is a multi-domain Roco protein, harbouring kinase and GTPase activity. In analogy with a bacterial homologue, LRRK2 was proposed to act as a GTPase activated by dimerization (GAD), while recent reports suggest LRRK2 to exist under a monomeric and dimeric form in vivo. It is however unknown how LRRK2 oligomerization is regulated. Here, we show that oligomerization of a homologous bacterial Roco protein depends on the nucleotide load. The protein is mainly dimeric in the nucleotide-free and GDP-bound states, while it forms monomers upon GTP binding, leading to a monomer-dimer cycle during GTP hydrolysis. An analogue of a PD-associated mutation stabilizes the dimer and decreases the GTPase activity. This work thus provides insights into the conformational cycle of Roco proteins and suggests a link between oligomerization and disease-associated mutations in LRRK2.

Contact author

• Egon Deyaert (VUB, Vrije Universiteit Brussel, Pleinlaan 2 1050 Brussel)

Models

Sample

• Sample: Name: Rab family protein CtRoc-COR GppNHp / Specimen concentration: 1.00-2.00
• Buffer: Name: 20 mM HEPES 150 mM NaCl 5 mM MgCl2 5% Glycerol 1 mM DTT 200µM GppNHp; pH: 7.5

Entity #605

Type: protein / Description: Rab family protein / Formula weight: 64.865 / Num. of mol.: 2
Source: Chlorobium tepidum (strain ATCC 49652 / DSM 12025 / NBRC 103806 / TLS)
References: UniProt: Q8KC98_CHL
Sequence:

MSYYHHHHHH DYDIPTTENL YFQGAMGSDN PLESPPPEIV KQGKEAVRQY FQSIEEARSK GEALVHLQEI KVHLIGDGMA GKTSLLKQLI GETFDPKESQ THGLNVVTKQ APNIKGLEND DELKECLFHF WDFGGQEIMH ASHQFFMTRS SVYMLLLDSR TDSNKHYWLR HIEKYGGKSP VIVVMNKIDE NPSYNIEQKK INERFPAIEN RFHRISCKNG DGVESIAKSL KSAVLHPDSI YGTPLAPSWI KVKEKLVEAT TAQRYLNRTE VEKICNDSGI TDPGERKTLL GYLNNLGIVL YFEALDLSEI YVLDPHWVTI GVYRIINSSK TKNGHLNTSA LGYILNEEQI RCDEYDPAKN NKFTYTLLEQ RYLLDIMKQF ELCYDEGKGL FIIPSNLPTQ IDNEPEITEG EPLRFIMKYD YLPSTIIPRL MIAMQHQILD RMQWRYGMVL KSQDHEGALA KVVAETKDST ITIAIQGEPR CKREYLSIIW YEIKKINANF TNLDVKEFIP LPGHPDELVE YKELLGLEKM GRDEYVSGKL EKVFSVSKML DSVISKEERN KER

Experimental information

• Beam: Instrument name: SOLEIL SWING / City: Saint-Aubin / 国: France / Type of source: X-ray synchrotronSynchrotron / Wavelength: 0.103 Å / Dist. spec. to detc.: 25 mm
• Detector: Name: AVIEX / Type: CCD

Scan

Title: Rab family protein CtRoc-COR GppNHp / Measurement date: Feb 1, 2017 / Exposure time: 0.75 sec. / Number of frames: 31 / Unit: 1/A /

	Min	Max
Q	0.004	0.6134

Distance distribution function P(R)

Sofotware P(R): GNOM 4.6 / Number of points: 475 /

	Min	Max
Q	0.01819	0.2875
P(R) point	26	500
R	0	105

Result

Type of curve: other /

	Experimental	Porod
MW	78 kDa	90 kDa
Volume	-	153.69 nm3

	P(R)	P(R) error	Guinier	Guinier error
Forward scattering, I0	0.06889	3.0E-5	0.069238	5.0E-5
Radius of gyration, Rg	3.331 nm	0.002	3.36 nm	0.003

	Min	Max
D	-	10.5
Guinier point	26	61