ArticleMethods in molecular biology (Clifton, N.J.)2025
Using GRAM Domain-Based Biosensors to Monitor Accessible Cholesterol Distribution in Live Cells.
Article in Methods in molecular biology (Clifton, N.J.), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.
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Who cites it
3 citing papers in PubMed.
- Cholesterol maintains the degradative capacity of lysosomes during clearance and recycling of dysfunctional mitochondria.Nature communications · 2026Article
- Live-Cell Imaging and Lipidomics of Low-Density Lipoprotein Containing Intrinsically Fluorescent Cholesteryl Esters.Analytical chemistry · 2026Article
- T cell cholesterol transport links intestinal immune responses to dietary lipid absorption.Science (New York, N.Y.) · 2025Article
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Authors and funding
4 authors.
Funding
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Abstract
Cholesterol is a vital component of cellular membranes, playing a crucial role in membrane integrity and cell signaling. It is unevenly distributed among different cellular compartments, with the highest enrichment in the plasma membrane (PM). Dysregulation of cellular cholesterol distribution is linked to various disorders, including cardiovascular and neurological diseases. Regulated transport of a specific pool of cholesterol, known as accessible cholesterol, is essential for maintaining cellular cholesterol distribution, yet visualization tools for accessible cholesterol in live cells are limited. Here, we introduce methods for visualizing accessible cholesterol using biosensors based on the GRAM domain of GRAMD1s/Asters (GRAMD1a, GRAMD1b, and GRAMD1c). We present two variants of the GRAM domain of GRAMD1b, GRAM-H, and GRAM-W, with different affinities for cholesterol, enabling the detection of both increase and decrease of accessible cholesterol in the PM. Additionally, we demonstrate the successful coupling of dimerization-dependent fluorescent protein (ddFP) with these biosensors to monitor accessible PM cholesterol levels in cell populations using flow cytometry. These GRAM domain-based biosensors complement existing tools, providing new insights into the molecular mechanisms underlying accessible cholesterol distribution.
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Registered trials
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