The Parkinson’s disease kinase LRRK2 autophosphorylates its GTPase domain at multiple sites

Greggio, Elisa; Taymans, Jean-Marc; Zhen, Yuejun; Ryder, John; Vancraenenbroeck, Renée; Beilina, Alexandra; Sun, Peng; Deng, Junpeng; Jaffe, Howard; Baekelandt, Veerle; Merchant, Kalpana; Cookson, Mark

doi:10.1016/j.bbrc.2009.08.163

Cited by 138 publications

(145 citation statements)

References 29 publications

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“…During the revision stage of this paper, Greggio et al reported on the identification of autophosphorylation sites within the ROC domain of LRRK2 (25), some of which (e.g., Thr1410, Thr1491) correspond to those determined in this present study.…”

Section: Acknowledgmentmentioning

confidence: 80%

Identification of the Autophosphorylation Sites of LRRK2

2009

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Parkinson's disease (PD) is a major adult-onset neurodegenerative disorder affecting the extrapyramidal motor system. A subset of patients develop PD as an autosomal dominant trait, of which PARK8 caused by mutations in the leucine-rich repeat kinase 2 (LRRK2) gene is highlighted because of its high frequency and clinicopathological similarity to sporadic PD. Previous studies have suggested that overactivation of LRRK2 caused by missense mutations leads to neuronal toxicity in PARK8, although the regulatory mechanism that governs the kinase activity of LRRK2 remains unknown. In this study, we expressed the carboxyl-half fragments of LRRK2 (DeltaN-LRRK2) that harbors the kinase as well as the ras-like (ROC) domains in Sf9 cells, subjected them to in vitro phosphorylation reaction, and analyzed the autophosphorylation by matrix assisted laser desorption/ionization- time of flight (MALDI-TOF) mass spectrometer. We identified Ser1403, Thr1404, Thr1410, Thr1491 located within the ROC domain, as well as Thr1967 and Thr1969 in the kinase domain, as the autophosphorylation sites. Substitution of Thr1967, an autophosphorylation site located within the kinase domain, to Ala caused a significant decrease in the kinase activity, implicating Thr1967 in the kinase activity of LRRK2. Phosphospecific antibodies to the autophosphorylation sites specifically recognized full-length LRRK2 subjected to in vitro phosphorylation reaction, indicating that the autophosphorylation takes place in holoproteins. Further analysis of autophosphorylation will clarify the mechanism of activation of LRRK2, as well as the pathomechanism of PD in relation to overactivation of LRRK2.

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Section: Acknowledgmentmentioning

confidence: 80%

Identification of the Autophosphorylation Sites of LRRK2

2009

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“…LRRK2 has been found to autophosphorylate > 20 serine and threonine residues in vitro 37, 50, 56, 57, 58, 59, 60, 61. The majority of the autophosphorylation sites reside in the ROC domain, with only a few in the COR and kinase domains (Fig.…”

Section: Importance Of Lrrk2 Autophosphorylation and Constitutive Phomentioning

confidence: 99%

Cellular processes associated with LRRK2 function and dysfunction

Wallings¹,

2015

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Mutations in the leucine‐rich repeat kinase 2 (LRRK2)‐encoding gene are the most common cause of monogenic Parkinson's disease. The identification of LRRK2 polymorphisms associated with increased risk for sporadic Parkinson's disease, as well as the observation that LRRK2‐Parkinson's disease has a pathological phenotype that is almost indistinguishable from the sporadic form of disease, suggested LRRK2 as the culprit to provide understanding for both familial and sporadic Parkinson's disease cases. LRRK2 is a large protein with both GTPase and kinase functions. Mutations segregating with Parkinson's disease reside within the enzymatic core of LRRK2, suggesting that modification of its activity impacts greatly on disease onset and progression. Although progress has been made since its discovery in 2004, there is still much to be understood regarding LRRK2′s physiological and neurotoxic properties. Unsurprisingly, given the presence of multiple enzymatic domains, LRRK2 has been associated with a diverse set of cellular functions and signalling pathways including mitochondrial function, vesicle trafficking together with endocytosis, retromer complex modulation and autophagy. This review discusses the state of current knowledge on the role of LRRK2 in health and disease with discussion of potential substrates of phosphorylation and functional partners with particular emphasis on signalling mechanisms. In addition, the use of immune cells in LRRK2 research and the role of oxidative stress as a regulator of LRRK2 activity and cellular function are also discussed.

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“…Reciprocal regulation of GTP binding by the kinase domain may also occur, as LRRK2 autophosphorylates several residues within the ROC domain. Phosphorylation of T1491 and T1503 residues apparently diminishes GTP binding [24][25][26][27][28][29], suggesting that LRRK2 GTPase may be functionally downstream of the kinase activity [30], although more complex situations may occur. These suggestions show that there is intramolecular communication between different LRRK2 domains, pointing out how complex the molecule is.…”

Section: Introductionmentioning

confidence: 99%

Heterogeneity of Leucine-Rich Repeat Kinase 2 Mutations: Genetics, Mechanisms and Therapeutic Implications

Rudenko

Cookson

2014

Neurotherapeutics

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Variation within and around the leucine-rich repeat kinase 2 (LRRK2) gene is associated with familial and sporadic Parkinson's disease (PD). Here, we discuss the prevalence of LRRK2 substitutions in different populations and their association with PD, as well as molecular and cellular mechanisms of pathologically relevant LRRK2 mutations. Kinase activation was proposed as a universal molecular mechanism for all pathogenic LRRK2 mutations, but later reports revealed heterogeneity in the effect of mutations on different activities of LRRK2. One mutation (G2019S) increases kinase activity, whereas mutations in the Ras of complex proteins (ROC)-C-terminus of ROC (COR) bidomain impair the GTPase function of LRRK2. Some risk factor variants, including G2385R in the WD40 domain, actually decrease the kinase activity of LRRK2. We suggest a model where LRRK2 mutations exert different molecular mechanisms but interfere with normal cellular function of LRRK2 at different levels of the same downstream pathway. Finally, we discuss the current state of therapeutic approaches for LRRK2-related PD.

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The Parkinson’s disease kinase LRRK2 autophosphorylates its GTPase domain at multiple sites

Cited by 138 publications

References 29 publications

Identification of the Autophosphorylation Sites of LRRK2

Identification of the Autophosphorylation Sites of LRRK2

Cellular processes associated with LRRK2 function and dysfunction

Heterogeneity of Leucine-Rich Repeat Kinase 2 Mutations: Genetics, Mechanisms and Therapeutic Implications

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