The Inhibitory Properties of a Novel, Selective LMTK3 Kinase Inhibitor

Agnarelli, Alessandro; Betrán, Andrea Lauer; Papakyriakou, Athanasios; Vella, Viviana; Samuels, Mark E.; Papanastasopoulos, Panagiotis; Giamas, Christina; Mancini, Erika J.; Stebbing, Justin; Spencer, John; Cilibrasi, Chiara; Ditsiou, Angeliki; Giamas, Georgios

doi:10.3390/ijms24010865

Cited by 5 publications

(3 citation statements)

References 40 publications

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“…Significantly high levels of phosphorylation were seen for serine 232 (S232), the phosphorylation of which has been shown to be critical for the catalytic activity of LMTK3. 22 , 23 , 35 In addition to S232, LMTK3 was phosphorylated on 12 other amino acids, but the significance of these covalent modifications for its activity, membrane trafficking, or stability is unknown.…”

Section: Resultsmentioning

confidence: 99%

The brain-specific kinase LMTK3 regulates neuronal excitability by decreasing KCC2-dependent neuronal Cl− extrusion

Cho,

Kontou,

Smalley

et al. 2024

iScience

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Section: Resultsmentioning

confidence: 99%

The brain-specific kinase LMTK3 regulates neuronal excitability by decreasing KCC2-dependent neuronal Cl− extrusion

Cho,

Kontou,

Smalley

et al. 2024

iScience

Self Cite

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“…Cell Death and Apoptosis: The assay was performed as previously described. [63,64] Cells were transfected with either siCTRL or siPANK4 as described above and subjected to drug treatments as specified in the corresponding figures and figure legends. After 96 h, cells were stained using the Muse Annexin V Dead Cell Kit according to the manufacturer's instructions (Luminex, #MCH100105).…”

Section: Methodsmentioning

confidence: 99%

Kinome‐Wide Synthetic Lethal Screen Identifies PANK4 as a Modulator of Temozolomide Resistance in Glioblastoma

Vella,

Ditsiou,

Chalari

et al. 2024

Advanced Science

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Temozolomide (TMZ) represents the cornerstone of therapy for glioblastoma (GBM). However, acquisition of resistance limits its therapeutic potential. The human kinome is an undisputable source of druggable targets, still, current knowledge remains confined to a limited fraction of it, with a multitude of under‐investigated proteins yet to be characterized. Here, following a kinome‐wide RNAi screen, pantothenate kinase 4 (PANK4) isuncovered as a modulator of TMZ resistance in GBM. Validation of PANK4 across various TMZ‐resistant GBM cell models, patient‐derived GBM cell lines, tissue samples, as well as in vivo studies, corroborates the potential translational significance of these findings. Moreover, PANK4 expression is induced during TMZ treatment, and its expression is associated with a worse clinical outcome. Furthermore, a Tandem Mass Tag (TMT)‐based quantitative proteomic approach, reveals that PANK4 abrogation leads to a significant downregulation of a host of proteins with central roles in cellular detoxification and cellular response to oxidative stress. More specifically, as cells undergo genotoxic stress during TMZ exposure, PANK4 depletion represents a crucial event that can lead to accumulation of intracellular reactive oxygen species (ROS) and subsequent cell death. Collectively, a previously unreported role for PANK4 in mediating therapeutic resistance to TMZ in GBM is unveiled.

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“…A range of biophysical techniques and cell-based assays were used to identify a highly selective small molecule inhibitor of lemur tyrosine kinase 3 (LMTK3), which is a dual specificity serine/threonine kinase with an oncogenic role in various tumour types and a viable therapeutic target. The analyses included use of MST to obtain binding curves for the new inhibitor C36 (KD = 1.87 ± 0.2 µM) and an existing inhibitor C28 (KD = 2.50 ± 0.4 µM) with human LMTK3 [56].…”

Section: Sensor Kinasesmentioning

confidence: 99%

Microscale Thermophoresis Analysis of Membrane Proteins

Patching

2024

Preprint

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Microscale thermophoresis (MST) is an analytical technique for measuring biomolecular interactions. It is based on the physical phenomenon that particles move within temperature gradients, which is affected by their size, charge, hydration shell and conformation. The MST sample must contain a fluorescent target molecule used to observe the movement of particles, and this can be titrated with an unlabelled binding partner for quantifying the interaction. MST is highly sensitive, using relatively small amounts of sample, and it has no limitations on the size of the target biomolecule, on the affinity of the interaction or on the composition of the buffer and other sample components. This makes MST ideally suited to characterising interactions with membrane proteins, which can be studied in cell lysates, native membranes, solubilised in detergents or reconstituted in lipids. The intrinsic aromatic residues of membrane proteins have been used as the fluorophore for MST (label-free MST) or membrane proteins have been labelled with a range of fluorescent dyes or conjugated with fluorescent proteins (labelled MST). The different types of membrane proteins that have had biomolecular interactions characterised by MST include the SARS-CoV-2 spike protein, GPCRs and other receptors, sensor kinases, ion channels, aquaporins, and transport proteins.

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The Inhibitory Properties of a Novel, Selective LMTK3 Kinase Inhibitor

Cited by 5 publications

References 40 publications

The brain-specific kinase LMTK3 regulates neuronal excitability by decreasing KCC2-dependent neuronal Cl− extrusion

The brain-specific kinase LMTK3 regulates neuronal excitability by decreasing KCC2-dependent neuronal Cl− extrusion

Kinome‐Wide Synthetic Lethal Screen Identifies PANK4 as a Modulator of Temozolomide Resistance in Glioblastoma

Microscale Thermophoresis Analysis of Membrane Proteins

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