scholarly journals Distinct profiles of LRRK2 activation and Rab GTPase phosphorylation in clinical samples from different PD cohorts

2021 ◽  
Author(s):  
Lilian Peptropoulou-Vathi ◽  
Athina M Simitsi ◽  
Politymi-Eleni Valkimadi ◽  
Maria Kedariti ◽  
Lampros Dimitrakopoulos ◽  
...  
Keyword(s):  
Outcome Measures ◽  
Kinase Activity ◽  
Disease Status ◽  
Clinical Samples ◽  
Rab Gtpase ◽  
Rab Gtpases ◽  
Comprehensive Picture ◽  
Lrrk2 Kinase ◽  
A53t Mutation ◽  
Lrrk2 Kinase Activity

Despite several advances in the field, pharmacodynamic outcome measures reflective of LRRK2 kinase activity in clinical biofluids remain urgently needed. A variety of targets and approaches have been utilized including assessments of LRRK2 itself (levels, phosphorylation), or its substrates (e.g. Rab10 or other Rab GTPases). We have previously shown that intrinsic kinase activity of LRRK2 isolated from PBMCs of G2019S carriers is elevated, irrespective of disease status. In the present study we find that phosphorylation of Rab10 is also elevated in G2019S carriers, but only those with PD. Additionally, phosphorylation of this substrate is also elevated in 2 separate idiopathic PD cohorts, but not in carriers of the A53T mutation in α-synuclein. In contrast, Rab29 phosphorylation was specifically reduced in urinary exosomes from A53T and idiopathic PD patients. Taken together, our findings highlight the need for the assessment of multiple complimentary targets for a more comprehensive picture of the disease.

scholarly journals Roles of lysosomotropic agents on LRRK2 activation and Rab10 phosphorylation

2020 ◽  
Author(s):  
Tomoki Kuwahara ◽  
Kai Funakawa ◽  
Tadayuki Komori ◽  
Maria Sakurai ◽  
Gen Yoshii ◽  
...  
Keyword(s):  
Kinase Activity ◽  
Autosomal Dominant ◽  
Rab Gtpases ◽  
The Novel ◽  
Causative Gene ◽  
Drug Induced ◽  
Extracellular Release ◽  
Enzymatic Activation ◽  
Lrrk2 Kinase ◽  
Lrrk2 Kinase Activity

AbstractLeucine-rich repeat kinase 2 (LRRK2), the major causative gene product of autosomal-dominant Parkinson’s disease, is a protein kinase that phosphorylates a subset of Rab GTPases. Since pathogenic LRRK2 mutations increase its ability to phosphorylate Rab GTPases, elucidating the mechanisms of how Rab phosphorylation is regulated by LRRK2 is of great importance. We have previously reported that chloroquine-induced lysosomal stress facilitates LRRK2 phosphorylation of Rab10 to maintain lysosomal homeostasis. Here we reveal that Rab10 phosphorylation by LRRK2 is potently stimulated by treatment of cells with a set of lysosome stressors and clinically used lysosomotropic drugs. These agents commonly promoted the formation of LRRK2-coated enlarged lysosomes and extracellular release of lysosomal enzyme cathepsin B, the latter being dependent on LRRK2 kinase activity. In contrast to the increase in Rab10 phosphorylation, treatment with lysosomotropic drugs did not increase the enzymatic activity of LRRK2, as monitored by its autophosphorylation at Ser1292 residue, but rather enhanced the molecular proximity between LRRK2 and its substrate Rab GTPases on the cytosolic surface of lysosomes. Lysosomotropic drug-induced upregulation of Rab10 phosphorylation was likely a downstream event of Rab29 (Rab7L1)-mediated enzymatic activation of LRRK2. These results suggest a regulated process of Rab10 phosphorylation by LRRK2 that is associated with lysosomal overload stress, and provide insights into the novel strategies to halt the aberrant upregulation of LRRK2 kinase activity.

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

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 The activities of LRRK2 and GCase are positively correlated in clinical biospecimens and experimental models of Parkinson’s disease

2021 ◽  
Author(s):  
Maria Kedariti ◽  
Emanuele Frattini ◽  
Pascale Baden ◽  
Susanna Cogo ◽  
Laura Civiero ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Kinase Activity ◽  
Experimental Models ◽  
Cellular Functions ◽  
Gene Encoding ◽  
Cellular Models ◽  
Lrrk2 Kinase ◽  
The Impact ◽  
Lrrk2 Kinase Activity

AbstractLRRK2 is a kinase involved in different cellular functions, including autophagy, endolysosomal pathways and vesicle trafficking. Mutations in LRRK2 cause autosomal dominant forms of Parkinson’s disease (PD). Heterozygous mutations in GBA1, the gene encoding the lysosomal enzyme glucocerebrosidase (GCase), are the most common genetic risk factors for PD. Moreover, GCase function is altered in idiopathic PD and in other genetic forms of the disease. Recent work suggests that LRRK2 kinase activity can regulate GCase function. However, both a positive and a negative correlation have been described. To gain insights into the impact of LRRK2 on GCase, we investigated GCase levels and activity in LRRK2 G2019S knockin mice, in clinical biospecimens from PD patients carrying this mutation and in patient-derived cellular models. In these models we found a positive correlation between the activities of LRRK2 and GCase, which was further confirmed in cell lines with genetic and pharmacological manipulation of LRRK2 kinase activity. Overall, our study indicates that LRRK2 kinase activity affects both the levels and the catalytic activity of GCase.

scholarly journals Crystal structure of the WD40 domain dimer of LRRK2

2019 ◽  
Vol 116 (5) ◽  
pp. 1579-1584 ◽  
Author(s):  
Pengfei Zhang ◽  
Ying Fan ◽  
Heng Ru ◽  
Li Wang ◽  
Venkat Giri Magupalli ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Kinase Activity ◽  
Kinase Inhibitors ◽  
Human Genetics ◽  
Kinase Domain ◽  
Rab Gtpases ◽  
Kinase Activation ◽  
Surface Patches ◽  
Wd40 Domain ◽  
Lrrk2 Kinase

Leucine-rich repeat kinase 2 (LRRK2) is a large multidomain protein with both a Ras of complex (ROC) domain and a kinase domain (KD) and, therefore, exhibits both GTPase and kinase activities. Human genetics studies have linked LRRK2 as a major genetic contributor to familial and sporadic Parkinson’s disease (PD), a neurodegenerative movement disorder that inflicts millions worldwide. The C-terminal region of LRRK2 is a Trp-Asp-40 (WD40) domain with poorly defined biological functions but has been implicated in microtubule interaction. Here, we present the crystal structure of the WD40 domain of human LRRK2 at 2.6-Å resolution, which reveals a seven-bladed WD40 fold. The structure displays a dimeric assembly in the crystal, which we further confirm by measurements in solution. We find that structure-based and PD-associated disease mutations in the WD40 domain including the common G2385R polymorphism mainly compromise dimer formation. Assessment of full-length LRRK2 kinase activity by measuring phosphorylation of Rab10, a member of the family of Rab GTPases known to be important kinase substrates of LRRK2, shows enhancement of kinase activity by several dimerization-defective mutants including G2385R, although dimerization impairment does not always result in kinase activation. Furthermore, mapping of phylogenetically conserved residues onto the WD40 domain structure reveals surface patches that may be important for additional functions of LRRK2. Collectively, our analyses provide insights for understanding the structures and functions of LRRK2 and suggest the potential utility of LRRK2 kinase inhibitors in treating PD patients with WD40 domain mutations.

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