Proteolysis of Rab32 by Salmonella GtgE induces an inactive GTPase conformation

Sergey Savitskiy; Rudolf Wachtel; Danial Pourjafar-Dehkordi; Hyun-Seo Kang; Vanessa Trauschke; Don C Lamb; Michael Sattler; Martin Zacharias; Aymelt Itzen

doi:10.1016/j.isci.2020.101940

Proteolysis of Rab32 by Salmonella GtgE induces an inactive GTPase conformation

Standard

Proteolysis of Rab32 by Salmonella GtgE induces an inactive GTPase conformation. / Savitskiy, Sergey; Wachtel, Rudolf; Pourjafar-Dehkordi, Danial; Kang, Hyun-Seo; Trauschke, Vanessa; Lamb, Don C; Sattler, Michael; Zacharias, Martin; Itzen, Aymelt.

In: ISCIENCE, Vol. 24, No. 1, 101940, 22.01.2021.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Savitskiy, S, Wachtel, R, Pourjafar-Dehkordi, D, Kang, H-S, Trauschke, V, Lamb, DC, Sattler, M, Zacharias, M & Itzen, A 2021, 'Proteolysis of Rab32 by Salmonella GtgE induces an inactive GTPase conformation', ISCIENCE, vol. 24, no. 1, 101940. https://doi.org/10.1016/j.isci.2020.101940

APA

Savitskiy, S., Wachtel, R., Pourjafar-Dehkordi, D., Kang, H-S., Trauschke, V., Lamb, D. C., Sattler, M., Zacharias, M., & Itzen, A. (2021). Proteolysis of Rab32 by Salmonella GtgE induces an inactive GTPase conformation. ISCIENCE, 24(1), [101940]. https://doi.org/10.1016/j.isci.2020.101940

Vancouver

Savitskiy S, Wachtel R, Pourjafar-Dehkordi D, Kang H-S, Trauschke V, Lamb DC et al. Proteolysis of Rab32 by Salmonella GtgE induces an inactive GTPase conformation. ISCIENCE. 2021 Jan 22;24(1). 101940. https://doi.org/10.1016/j.isci.2020.101940

Bibtex

@article{2db6de9c0d914f56ad8816e85a12cbcb,

title = "Proteolysis of Rab32 by Salmonella GtgE induces an inactive GTPase conformation",

abstract = "Rab GTPases are central regulators of intracellular vesicular trafficking. They are frequently targeted by bacterial pathogens through post-translational modifications. Salmonella typhimurium secretes the cysteine protease GtgE during infection, leading to a regioselective proteolytic cleavage of the regulatory switch I loop in the small GTPases of the Rab32 subfamily. Here, using a combination of biochemical methods, molecular dynamics simulations, NMR spectroscopy, and single-pair F{\"o}rster resonance energy transfer, we demonstrate that the cleavage of Rab32 causes a local increase of conformational flexibility in both switch regions. Cleaved Rab32 maintains its ability to interact with the GDP dissociation inhibitor (GDI). Interestingly, the Rab32 cleavage enables GDI binding also with an active GTP-bound Rab32 in vitro. Furthermore, the Rab32 proteolysis provokes disturbance in the interaction with its downstream effector VARP. Thus, the proteolysis of Rab32 is not a globally degradative mechanism but affects various biochemical and structural properties of the GTPase in a diverse manner.",

author = "Sergey Savitskiy and Rudolf Wachtel and Danial Pourjafar-Dehkordi and Hyun-Seo Kang and Vanessa Trauschke and Lamb, {Don C} and Michael Sattler and Martin Zacharias and Aymelt Itzen",

note = "{\textcopyright} 2020 The Author(s).",

year = "2021",

month = jan,

day = "22",

doi = "10.1016/j.isci.2020.101940",

language = "English",

volume = "24",

journal = "ISCIENCE",

issn = "2589-0042",

publisher = "Elsevier Inc.",

number = "1",

}

RIS

TY - JOUR

T1 - Proteolysis of Rab32 by Salmonella GtgE induces an inactive GTPase conformation

AU - Savitskiy, Sergey

AU - Wachtel, Rudolf

AU - Pourjafar-Dehkordi, Danial

AU - Kang, Hyun-Seo

AU - Trauschke, Vanessa

AU - Lamb, Don C

AU - Sattler, Michael

AU - Zacharias, Martin

AU - Itzen, Aymelt

PY - 2021/1/22

Y1 - 2021/1/22

N2 - Rab GTPases are central regulators of intracellular vesicular trafficking. They are frequently targeted by bacterial pathogens through post-translational modifications. Salmonella typhimurium secretes the cysteine protease GtgE during infection, leading to a regioselective proteolytic cleavage of the regulatory switch I loop in the small GTPases of the Rab32 subfamily. Here, using a combination of biochemical methods, molecular dynamics simulations, NMR spectroscopy, and single-pair Förster resonance energy transfer, we demonstrate that the cleavage of Rab32 causes a local increase of conformational flexibility in both switch regions. Cleaved Rab32 maintains its ability to interact with the GDP dissociation inhibitor (GDI). Interestingly, the Rab32 cleavage enables GDI binding also with an active GTP-bound Rab32 in vitro. Furthermore, the Rab32 proteolysis provokes disturbance in the interaction with its downstream effector VARP. Thus, the proteolysis of Rab32 is not a globally degradative mechanism but affects various biochemical and structural properties of the GTPase in a diverse manner.

AB - Rab GTPases are central regulators of intracellular vesicular trafficking. They are frequently targeted by bacterial pathogens through post-translational modifications. Salmonella typhimurium secretes the cysteine protease GtgE during infection, leading to a regioselective proteolytic cleavage of the regulatory switch I loop in the small GTPases of the Rab32 subfamily. Here, using a combination of biochemical methods, molecular dynamics simulations, NMR spectroscopy, and single-pair Förster resonance energy transfer, we demonstrate that the cleavage of Rab32 causes a local increase of conformational flexibility in both switch regions. Cleaved Rab32 maintains its ability to interact with the GDP dissociation inhibitor (GDI). Interestingly, the Rab32 cleavage enables GDI binding also with an active GTP-bound Rab32 in vitro. Furthermore, the Rab32 proteolysis provokes disturbance in the interaction with its downstream effector VARP. Thus, the proteolysis of Rab32 is not a globally degradative mechanism but affects various biochemical and structural properties of the GTPase in a diverse manner.

U2 - 10.1016/j.isci.2020.101940

DO - 10.1016/j.isci.2020.101940

M3 - SCORING: Journal article

C2 - 33426511

VL - 24

JO - ISCIENCE

JF - ISCIENCE

SN - 2589-0042

IS - 1

M1 - 101940

ER -