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Allosteric modulation of the GTPase activity of a bacterial LRRK2 homolog by conformation-specific Nanobodies. / Leemans, Margaux; Galicia, Christian; Deyaert, Egon; Daems, Elise; Krause, Linda; Paesmans, Jone; Pardon, Els; Steyaert, Jan; Kortholt, Arjan; Sobott, Frank; Klostermeier, Dagmar; Versees, Wim.

In: Biochemical Journal, Vol. 477, No. 7, 04.2020, p. 1203-1218.

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Leemans, Margaux ; Galicia, Christian ; Deyaert, Egon ; Daems, Elise ; Krause, Linda ; Paesmans, Jone ; Pardon, Els ; Steyaert, Jan ; Kortholt, Arjan ; Sobott, Frank ; Klostermeier, Dagmar ; Versees, Wim. / Allosteric modulation of the GTPase activity of a bacterial LRRK2 homolog by conformation-specific Nanobodies. In: Biochemical Journal. 2020 ; Vol. 477, No. 7. pp. 1203-1218.

BibTeX

@article{5cd8082b6e2043e8950bd5ac9a4a255a,
title = "Allosteric modulation of the GTPase activity of a bacterial LRRK2 homolog by conformation-specific Nanobodies",
abstract = "Mutations in the Parkinson's disease (PD)-associated protein leucine-rich repeat kinase 2 (LRRK2) commonly lead to a reduction of GTPase activity and increase in kinase activity. Therefore, strategies for drug development have mainly been focusing on the design of LRRK2 kinase inhibitors. We recently showed that the central RocCOR domains (Roc: Ras of complex proteins; COR: C-terminal of Roc) of a bacterial LRRK2 homolog cycle between a dimeric and monomeric form concomitant with GTP binding and hydrolysis. PD-associated mutations can slow down GTP hydrolysis by stabilizing the protein in its dimeric form. Here, we report the identification of two Nanobodies (NbRoco1 and NbRoco2) that bind the bacterial Roco protein (CtRoco) in a conformation-specific way, with a preference for the GTP-bound state. NbRoco1 considerably increases the GTP turnover rate of CtRoco and reverts the decrease in GTPase activity caused by a PD-analogous mutation. We show that NbRoco1 exerts its effect by allosterically interfering with the CtRoco dimer-monomer cycle through the destabilization of the dimeric form. Hence, we provide the first proof of principle that allosteric modulation of the RocCOR dimer-monomer cycle can alter its GTPase activity, which might present a potential novel strategy to overcome the effect of LRRK2 PD mutations.",
author = "Margaux Leemans and Christian Galicia and Egon Deyaert and Elise Daems and Linda Krause and Jone Paesmans and Els Pardon and Jan Steyaert and Arjan Kortholt and Frank Sobott and Dagmar Klostermeier and Wim Versees",
year = "2020",
month = "4",
doi = "10.1042/BCJ20190843",
language = "English",
volume = "477",
pages = "1203--1218",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "7",

}

RIS

TY - JOUR

T1 - Allosteric modulation of the GTPase activity of a bacterial LRRK2 homolog by conformation-specific Nanobodies

AU - Leemans, Margaux

AU - Galicia, Christian

AU - Deyaert, Egon

AU - Daems, Elise

AU - Krause, Linda

AU - Paesmans, Jone

AU - Pardon, Els

AU - Steyaert, Jan

AU - Kortholt, Arjan

AU - Sobott, Frank

AU - Klostermeier, Dagmar

AU - Versees, Wim

PY - 2020/4

Y1 - 2020/4

N2 - Mutations in the Parkinson's disease (PD)-associated protein leucine-rich repeat kinase 2 (LRRK2) commonly lead to a reduction of GTPase activity and increase in kinase activity. Therefore, strategies for drug development have mainly been focusing on the design of LRRK2 kinase inhibitors. We recently showed that the central RocCOR domains (Roc: Ras of complex proteins; COR: C-terminal of Roc) of a bacterial LRRK2 homolog cycle between a dimeric and monomeric form concomitant with GTP binding and hydrolysis. PD-associated mutations can slow down GTP hydrolysis by stabilizing the protein in its dimeric form. Here, we report the identification of two Nanobodies (NbRoco1 and NbRoco2) that bind the bacterial Roco protein (CtRoco) in a conformation-specific way, with a preference for the GTP-bound state. NbRoco1 considerably increases the GTP turnover rate of CtRoco and reverts the decrease in GTPase activity caused by a PD-analogous mutation. We show that NbRoco1 exerts its effect by allosterically interfering with the CtRoco dimer-monomer cycle through the destabilization of the dimeric form. Hence, we provide the first proof of principle that allosteric modulation of the RocCOR dimer-monomer cycle can alter its GTPase activity, which might present a potential novel strategy to overcome the effect of LRRK2 PD mutations.

AB - Mutations in the Parkinson's disease (PD)-associated protein leucine-rich repeat kinase 2 (LRRK2) commonly lead to a reduction of GTPase activity and increase in kinase activity. Therefore, strategies for drug development have mainly been focusing on the design of LRRK2 kinase inhibitors. We recently showed that the central RocCOR domains (Roc: Ras of complex proteins; COR: C-terminal of Roc) of a bacterial LRRK2 homolog cycle between a dimeric and monomeric form concomitant with GTP binding and hydrolysis. PD-associated mutations can slow down GTP hydrolysis by stabilizing the protein in its dimeric form. Here, we report the identification of two Nanobodies (NbRoco1 and NbRoco2) that bind the bacterial Roco protein (CtRoco) in a conformation-specific way, with a preference for the GTP-bound state. NbRoco1 considerably increases the GTP turnover rate of CtRoco and reverts the decrease in GTPase activity caused by a PD-analogous mutation. We show that NbRoco1 exerts its effect by allosterically interfering with the CtRoco dimer-monomer cycle through the destabilization of the dimeric form. Hence, we provide the first proof of principle that allosteric modulation of the RocCOR dimer-monomer cycle can alter its GTPase activity, which might present a potential novel strategy to overcome the effect of LRRK2 PD mutations.

U2 - 10.1042/BCJ20190843

DO - 10.1042/BCJ20190843

M3 - Article

VL - 477

SP - 1203

EP - 1218

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 7

ER -

ID: 51826754