Pharmacological inhibition of LRRK2 cellular phosphorylation sites provides insight into LRRK2 biology

2012 ◽  
Vol 40 (5) ◽  
pp. 1158-1162 ◽  
Author(s):  
Jing Zhao ◽  
Spencer B. Hermanson ◽  
Coby B. Carlson ◽  
Steven M. Riddle ◽  
Kurt W. Vogel ◽  
...  
Keyword(s):  
High Throughput Screening ◽  
Kinase Activity ◽  
Phosphorylation Sites ◽  
Dependent Manner ◽  
Neuroprotective Agents ◽  
Leucine Rich Repeat ◽  
Lrrk2 Kinase ◽  
Activity Dependent ◽  
Lrrk2 Kinase Activity ◽  
Insight Into

Mutations in LRRK2 (leucine-rich repeat kinase 2) have been linked to inherited forms of PD (Parkinson's disease). Substantial pre-clinical research and drug discovery efforts have focused on LRRK2 with the hope that small-molecule inhibitors of the enzyme may be valuable for the treatment or prevention of the onset of PD. The pathway to develop therapeutic or neuroprotective agents based on LRRK2 function (i.e. kinase activity) has been facilitated by the development of both biochemical and cell-based assays for LRRK2. LRRK2 is phosphorylated on Ser910, Ser935, Ser955 and Ser973 in the N-terminal domain of the enzyme, and these sites of phosphorylation are likely to be regulated by upstream enzymes in an LRRK2 kinase-activity-dependent manner. Knowledge of these phosphorylation sites and their regulation can be adapted to high-throughput-screening-amenable platforms. The present review describes the utilization of LRRK2 phosphorylation as indicators of enzyme inhibition, as well as how such assays can be used to deconvolute the pathways in which LRRK2 plays a role.

Phosphorylation of LRRK2: from kinase to substrate

2012 ◽  
Vol 40 (5) ◽  
pp. 1102-1110 ◽  
Author(s):  
Evy Lobbestael ◽  
Veerle Baekelandt ◽  
Jean-Marc Taymans
Keyword(s):  
Kinase Activity ◽  
Pathological Process ◽  
Rapid Decrease ◽  
Leucine Rich Repeat ◽  
Phosphorylated Protein ◽  
Pathogenic Variants ◽  
Disease Protein ◽  
Lrrk2 Kinase ◽  
Disease Modifying ◽  
Lrrk2 Kinase Activity

The PD (Parkinson's disease) protein LRRK2 (leucine-rich repeat kinase 2) occurs in cells as a highly phosphorylated protein, with the majority of phosphosites clustering in the region between the ankyrin repeat and leucine-rich repeat domains. The observation that several pathogenic variants of LRRK2 display strongly reduced cellular phosphorylation suggests that phosphorylation of LRRK2 is involved in the PD pathological process. Furthermore, treatment of cells with inhibitors of LRRK2 kinase activity, which are currently considered as potential disease-modifying therapeutics for PD, leads to a rapid decrease in the phosphorylation levels of LRRK2. For these reasons, understanding the cellular role and regulation of LRRK2 as a kinase and as a substrate has become the focus of intense investigation. In the present review, we discuss what is currently known about the cellular phosphorylation of LRRK2 and how this relates to its function and dysfunction.

scholarly journals P62/SQSTM1 is a novel leucine-rich repeat kinase 2 (LRRK2) substrate that enhances neuronal toxicity

2018 ◽  
Vol 475 (7) ◽  
pp. 1271-1293 ◽  
Author(s):  
Alexia F. Kalogeropulou ◽  
Jing Zhao ◽  
Marc F. Bolliger ◽  
Anna Memou ◽  
Shreya Narasimha ◽  
...  
Keyword(s):  
Kinase Activity ◽  
Rab Proteins ◽  
Missense Mutations ◽  
Leucine Rich Repeat ◽  
Lrrk2 Gene ◽  
Ubiquitin Binding Domain ◽  
Lrrk2 Kinase ◽  
Lrrk2 Kinase Activity ◽  
In Cells

Autosomal-dominant, missense mutations in the leucine-rich repeat protein kinase 2 (LRRK2) gene are the most common genetic predisposition to develop Parkinson's disease (PD). LRRK2 kinase activity is increased in several pathogenic mutations (N1437H, R1441C/G/H, Y1699C, G2019S), implicating hyperphosphorylation of a substrate in the pathogenesis of the disease. Identification of the downstream targets of LRRK2 is a crucial endeavor in the field to understand LRRK2 pathway dysfunction in the disease. We have identified the signaling adapter protein p62/SQSTM1 as a novel endogenous interacting partner and a substrate of LRRK2. Using mass spectrometry and phospho-specific antibodies, we found that LRRK2 phosphorylates p62 on Thr138 in vitro and in cells. We found that the pathogenic LRRK2 PD-associated mutations (N1437H, R1441C/G/H, Y1699C, G2019S) increase phosphorylation of p62 similar to previously reported substrate Rab proteins. Notably, we found that the pathogenic I2020T mutation and the risk factor mutation G2385R displayed decreased phosphorylation of p62. p62 phosphorylation by LRRK2 is blocked by treatment with selective LRRK2 inhibitors in cells. We also found that the amino-terminus of LRRK2 is crucial for optimal phosphorylation of Rab7L1 and p62 in cells. LRRK2 phosphorylation of Thr138 is dependent on a p62 functional ubiquitin-binding domain at its carboxy-terminus. Co-expression of p62 with LRRK2 G2019S increases the neurotoxicity of this mutation in a manner dependent on Thr138. p62 is an additional novel substrate of LRRK2 that regulates its toxic biology, reveals novel signaling nodes and can be used as a pharmacodynamic marker for LRRK2 kinase activity.

scholarly journals Dependence of Leucine-rich Repeat Kinase 2 (LRRK2) Kinase Activity on Dimerization

2009 ◽  
Vol 284 (52) ◽  
pp. 36346-36356 ◽  
Author(s):  
Saurabh Sen ◽  
Philip J. Webber ◽  
Andrew B. West
Keyword(s):  
Kinase Activity ◽  
Leucine Rich Repeat ◽  
Lrrk2 Kinase ◽  
Lrrk2 Kinase Activity

scholarly journals Allosteric inhibition of LRRK2, where are we now

2020 ◽  
Vol 48 (5) ◽  
pp. 2185-2194
Author(s):  
Ahmed Soliman ◽  
Fatma Nihan Cankara ◽  
Arjan Kortholt
Keyword(s):  
Recent Progress ◽  
Kinase Activity ◽  
Kinase Inhibitors ◽  
High Selectivity ◽  
Gtpase Activity ◽  
Leucine Rich Repeat ◽  
Allosteric Inhibitors ◽  
Lrrk2 Kinase ◽  
Lrrk2 Kinase Activity

Parkinson's disease (PD) is the second most common neurodegenerative disease. In recent years, it has been shown that leucine-rich repeat kinase 2 (LRRK2) has a crucial function in both familial and sporadic forms of PD. LRRK2 pathogenic mutations are thought to result in an increase in LRRK2 kinase activity. Thus, inhibiting LRRK2 kinase activity has become a main therapeutic target. Many compounds capable of inhibiting LRRK2 kinase activity with high selectivity and brain availability have been described. However, the safety of long-term use of these ATP-competitive LRRK2 kinase inhibitors has been challenged by several studies. Therefore, alternative ways of targeting LRRK2 activity will have a great benefit. In this review, we discuss the recent progress in the development of allosteric inhibitors of LRRK2, mainly via interfering with GTPase activity, and propose potential new intra and interprotein interactions targets that can lead to open doors toward new therapeutics.

scholarly journals Increased LRRK2 kinase activity alters neuronal autophagy by disrupting the axonal transport of autophagosomes

2021 ◽  
Author(s):  
C. Alexander Boecker ◽  
Juliet Goldsmith ◽  
Dan Dou ◽  
Gregory G. Cajka ◽  
Erika L.F. Holzbaur
Keyword(s):  
Axonal Transport ◽  
Kinase Activity ◽  
Lrrk2 Kinase ◽  
Lrrk2 Kinase Activity ◽  
Neuronal Autophagy

scholarly journals Vitamin B12 modulates Parkinson’s disease LRRK2 kinase activity through allosteric regulation and confers neuroprotection

Cell Research ◽  
2019 ◽  
Vol 29 (4) ◽  
pp. 313-329 ◽  
Author(s):  
Adam Schaffner ◽  
Xianting Li ◽  
Yacob Gomez-Llorente ◽  
Emmanouela Leandrou ◽  
Anna Memou ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Vitamin B12 ◽  
Kinase Activity ◽  
Allosteric Regulation ◽  
Lrrk2 Kinase ◽  
Lrrk2 Kinase Activity

Ser1292 Autophosphorylation Is an Indicator of LRRK2 Kinase Activity and Contributes to the Cellular Effects of PD Mutations

2012 ◽  
Vol 4 (164) ◽  
pp. 164ra161-164ra161 ◽  
Author(s):  
Z. Sheng ◽  
S. Zhang ◽  
D. Bustos ◽  
T. Kleinheinz ◽  
C. E. Le Pichon ◽  
...  
Keyword(s):  
Kinase Activity ◽  
Cellular Effects ◽  
Lrrk2 Kinase ◽  
Lrrk2 Kinase Activity

Measurement of LRRK2 Kinase Activity by Proximity Ligation Assay

BIO-PROTOCOL ◽  
2021 ◽  
Vol 11 (17) ◽  
Author(s):  
Matthew Keeney ◽  
Eric Hoffman ◽  
J. Greenamyre ◽  
Roberto Di Maio
Keyword(s):  
Kinase Activity ◽  
Proximity Ligation Assay ◽  
Proximity Ligation ◽  
Lrrk2 Kinase ◽  
Lrrk2 Kinase Activity

scholarly journals ZBP1 induces inflammatory signaling via RIPK3 and promotes SARS-CoV-2-induced cytokine expression

2021 ◽  
Author(s):  
Ruoshi Peng ◽  
Xuan Wang-Kan ◽  
Manja Idorn ◽  
Felix Y Zhou ◽  
Susana L Orozco ◽  
...  
Keyword(s):  
Cell Death ◽  
Signaling Pathway ◽  
Kinase Activity ◽  
Caspase 8 ◽  
Dependent Manner ◽  
Innate Immune Responses ◽  
Inflammatory Signaling ◽  
Antiviral Responses ◽  
Nucleic Acid Sensor ◽  
Activity Dependent

AbstractCOVID-19 caused by the SARS-CoV-2 virus remains a threat to global health. The disease severity is mediated by cell death and inflammation, which regulate both the antiviral and the pathological innate immune responses. ZBP1, an interferon-induced cytosolic nucleic acid sensor, facilitates antiviral responses via RIPK3. Although ZBP1-mediated cell death is widely described, whether and how it promotes inflammatory signaling is unclear. Here, we report a ZBP1-induced inflammatory signaling pathway that depends on ubiquitination and RIPK3’s scaffolding ability independently of cell death. In human cells, ZBP1 associates with RIPK1 and RIPK3 as well as ubiquitin ligases cIAP1 and LUBAC. RIPK1 and ZBP1 are ubiquitinated to promote TAK1- and IKK-mediated inflammatory signaling. Additionally, RIPK1 recruits the p43/41-caspase-8-p43-FLIP heterodimer to suppress RIPK3 kinase activity, which otherwise promotes inflammatory signaling in a kinase activity-dependent manner. Lastly, we show that ZBP1 contributes to SARS-CoV-2-induced cytokine production. Taken together, we describe a ZBP1-RIPK1-RIPK3-mediated inflammatory signaling pathway relayed by the scaffolding role of RIPKs and regulated by caspase-8. Our results suggest the ZBP1 pathway contributes to inflammation in response to SARS-CoV-2 infection.

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