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PreprintCited by 5Parkinson's Disease Mechanisms and TreatmentsProtein Kinase Regulation and GTPase SignalingProtein Structure and Dynamics

The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.

Development of LRRK2 designed ankyrin-repeat proteins

Open access
Dederer, Verena, Murillo, Marta Sanz, Karasmanis, Eva P, Hatch, Kathryn S, Chatterjee, Deep, Preuss, Franziska, Abdul Azeez, Kamal R, Nguyen, Landon Vu, Galicia, Christian, Dreier, Birgit, Plückthun, Andreas, Versees, Wim, Mathea, Sebastian, Leschziner, Andres E, Reck-Peterson, Samara L, Knapp, Stefan·2023·Journal of Biological Chemistry
DOI 10.1101/2023.11.14.567123PMC PMC11284679

Preprint — published as 10.1016/j.jbc.2024.107469

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Abstract

Leucine rich repeat kinase 2 (LRRK2) is a large multidomain protein containing two catalytic domains, a kinase and a GTPase, as well protein interactions domains, including a WD40 domain. The association of increased LRRK2 kinase activity with both the familial and sporadic forms of Parkinson’s disease (PD) has led to intense interest in determining its cellular function. However, small molecule probes that can bind to LRRK2 and report on or affect its activity are needed. Here, we identified a series of high-affinity LRRK2-binding designed ankyrin-repeat proteins (DARPins). One of these DARPins (E11) bound to the LRRK2 WD40 domain with high affinity. LRRK2 bound to DARPin E11 showed improved behavior on cryo-EM grids, resulting in higher resolution LRRK2 structures. DARPin E11 did not affect the catalytic activity of a truncated form of LRRK2 in vitro but decreased the phosphorylation of Rab8A, a LRRK2 substrate, in cells. We also found that DARPin E11 disrupts the formation of microtubule-associated LRRK2 filaments in cells, which are known to require WD40-based dimerization. Thus, DARPin E11 is a new tool to explore the function and dysfunction of LRRK2 and guide the development of LRRK2 kinase inhibitors that target the WD40 domain instead of the kinase.

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Metadata

Authors
Dederer, V., Murillo, M. S., Karasmanis, E. P., Hatch, K. S., Chatterjee, D., Preuss, F., Abdul Azeez, K. R., Nguyen, L. V., Galicia, C., Dreier, B., Plückthun, A., Versees, W., Mathea, S., Leschziner, A. E., Reck-Peterson, S. L., & Knapp, S.
Affiliations
  • Goethe University Frankfurt (DE)
  • University of California San Diego (US)
  • Vrije Universiteit Brussel (BE)
  • University of Zurich (CH)
Corresponding author
Knapp, Stefan
ORCID
Venue
Journal of Biological Chemistry
Publisher
openRxiv
Published
2023-11-15
Volume
300
Issue
7
Pages
107469-107469
ISSN
0021-9258 (ISSN-L)
DOI
10.1101/2023.11.14.567123
PMC
PMC11284679
OpenAlex
W4399566509
Cited by
5
References
1
Citations by year
2026: 3 · 2025: 1 · 2023: 1
FWCI
1.06
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • eScholarship (California Digital Library) (submittedVersion, cc-by)
  • VUBIR (Vrije Universiteit Brussel) (submittedVersion)
  • Publication Server of Goethe University Frankfurt am Main (Goethe University Frankfurt) (submittedVersion, cc-by)
Funders
  • Howard Hughes Medical Institute
  • Michael J. Fox Foundation for Parkinson's Research
  • Pfizer
  • Bayer (875510)
  • McGill University
  • Genome Canada (875510)
  • European Federation of Pharmaceutical Industries and Associations (875510)
  • Aligning Science Across Parkinson’s
  • Innovative Health Initiative
  • Innovative Medicines Initiative
  • Genentech (875510)
  • Structural Genomics Consortium
Topics
  • Health Sciences › Medicine › Clinical Neurology › Parkinson's Disease Mechanisms and Treatments
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
Keywords
LRRK2, Parkinson's disease, DARPin, WD40 domain, protein-protein binding, kinase inhibitors, chemical probes, cryo-EM structure
MeSH terms
Leucine-Rich Repeat Serine-Threonine Protein Kinase-2, Humans, Parkinson Disease, Phosphorylation, Protein Binding, Ankyrin Repeat, Cryoelectron Microscopy, rab GTP-Binding Proteins, HEK293 Cells

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