The Annual Meeting of the American Society for Neurochemistry

We hope to meet you at American Society for Neurochemistry in April.

Time:      April 14 – 18, 2024

Venue:    Portland

From Sophion, you can meet Daniel Sauter who is ready for a chat about ion channel and automated patch clamping.

During the meeting, Daniel Sauter, will be presenting a poster titled: Development and validation of TRPML1 assays on automated patch clamp platforms

Abstract:

TRPML1 is a cation-selective ion channel that is ubiquitously expressed in lysosomes and late endosomes. TRPML1 is intimately involved in the regulation of lysosomal Ca2+ homeostasis and is consequently a critical component of autophagy and lysosomal biogenesis. Malfunctions of these processes are linked to various neurodegenerative disorders, including Alzheimer`s and Parkinson`s disease. The relevance of TRPML1 in neurodegenerative disorders is further validated by a loss-of-function mutation that was shown to cause the neurodegenerative lysosomal storage disease mucolipidosis type IV (MLIV).

The patch clamp technique remains the gold standard for studying ion channels as recordings provide a direct measure of the protein’s activity. Whilst powerful, the conventional patch clamp method requires highly trained scientists and allows only a low throughput. Automated patch clamp systems have evolved to overcome these limitations, and these platforms are therefore well-suited to test large numbers of compounds in a short time to support ion channel drug development programs.

Here, an automated patch clamp assay for TRPML1 recombinantly expressed in HEK293 cells is presented. To achieve plasma membrane expression, the two dileucine motifs (15LL and 577LL) of mouse TRPML1 responsible for lysosomal targeting were replaced with alanines to allow trafficking to the plasma membrane. Biophysical characterization of the channel in the whole-cell configuration was in good agreement with the literature. Application of the tool compound ML-SA5 at pH4.6 resulted in a marked increase in current amplitude with EC50 = 3.4 µM.

In conclusion, the developed assay allows to record a large number of compounds in a short time span to identify novel modulators of the TRPML1 channel.

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