Illuminating understudied kinases: a generalizable biosensor development method applied to protein kinase N DOI Creative Commons
Julius Bogomolovas, Ju Chen

Communications Biology, Journal Year: 2025, Volume and Issue: 8(1)

Published: Jan. 22, 2025

Abstract Protein kinases play crucial roles in regulating cellular processes, making real-time visualization of their activity essential for understanding signaling dynamics. While genetically encoded fluorescent biosensors have emerged as powerful tools studying kinase activity, development many remains challenging due to the lack suitable substrate peptides. Here, we present a novel approach identifying peptide substrates and demonstrate its effectiveness by developing biosensor Kinase N (PKN) activity. Our method identified new PKN that optimized use design. The resulting shows specificity family can detect both overexpressed endogenous live cells. Importantly, our revealed sustained basal PKN2 at plasma membrane, it hotspot. This work not only provides valuable tool but also demonstrates promising strategy other understudied kinases, potentially expanding ability monitor across human kinome.

Language: Английский

Fasudil inhibits α-synuclein aggregation through ROCK-inhibition-mediated mechanisms DOI Creative Commons
Lucia Lage, Ana I. Rodríguez‐Pérez, José L. Labandeira‐García

et al.

Neurotherapeutics, Journal Year: 2025, Volume and Issue: unknown, P. e00544 - e00544

Published: Feb. 1, 2025

ROCK inhibitors such as fasudil protected against dopaminergic degeneration and other neurodegenerative processes in several experimental models through inhibition of neuroinflammation activation survival signaling pathways, clinical trials have been initiated. More recently, has suggested to inhibit α-synuclein aggregation. However, this is controversial, particularly if it a consequence direct binding the molecule α-synuclein. We studied mechanisms involved effects on aggregation using α-synuclein-T/V5-synphilin-1 model. Molecule-molecule interactions were real time quaking inducing conversion (RT-QuiC). Fasudil decreased number cells with inclusions size neurons glial cells, inhibited microglial endocytosis aggregates. These changes not due protein expression or phosphorylation related rather than interaction α-synuclein, confirmed second inhibitor (Y27632) gene silencing. observed that downregulates factors are known promote NADPH-oxidase-derived oxidative stress, intracellular calcium increase, endocytosis, promotes autophagy. The present results support useful drug Parkinson's disease progression. In addition reported neuroprotective properties, inhibits aggregates, which enhances inflammatory response. mostly inhibition, we shown two structurally different knockdown data, further supported by RT-QuiC.

Language: Английский

Citations

1
Illuminating understudied kinases: a generalizable biosensor development method applied to protein kinase N DOI Creative Commons
Julius Bogomolovas, Ju Chen

Communications Biology, Journal Year: 2025, Volume and Issue: 8(1)

Published: Jan. 22, 2025

Abstract Protein kinases play crucial roles in regulating cellular processes, making real-time visualization of their activity essential for understanding signaling dynamics. While genetically encoded fluorescent biosensors have emerged as powerful tools studying kinase activity, development many remains challenging due to the lack suitable substrate peptides. Here, we present a novel approach identifying peptide substrates and demonstrate its effectiveness by developing biosensor Kinase N (PKN) activity. Our method identified new PKN that optimized use design. The resulting shows specificity family can detect both overexpressed endogenous live cells. Importantly, our revealed sustained basal PKN2 at plasma membrane, it hotspot. This work not only provides valuable tool but also demonstrates promising strategy other understudied kinases, potentially expanding ability monitor across human kinome.

Language: Английский

Citations

0