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Journal / Biochemistry / A High-Throughput Luciferase Assay to Evaluate…
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Biochemistry

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JoVE Journal Biochemistry
A High-Throughput Luciferase Assay to Evaluate Proteolysis of the Single-Turnover Protease PCSK9
 

A High-Throughput Luciferase Assay to Evaluate Proteolysis of the Single-Turnover Protease PCSK9

Article DOI: 10.3791/58265-v • 08:15 min • August 28th, 2018
August 28th, 2018 •
1, 2, 2
1Division of Cardiology, Department of Medicine, Zuckerberg San Francisco General and University of California San Francisco, 2Department of Cellular and Molecular Pharmacology and Howard Hughes Medical Institute, University of California San Francisco

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Summary
Automatic Translation

This protocol presents a method to evaluate the proteolytic activity of an intrinsically low-activity, single turnover protease in a cellular context. Specifically, this method is applied to evaluate the proteolytic activity of PCSK9, a key driver of lipid metabolism whose proteolytic activity is required for its ultimate hypercholesterolemic function.

Tags

Luciferase Assay Proteolysis Single-turnover Protease PCSK9 Drugs Genetic Changes Biochemical Event Atherosclerotic Heart Disease Serum Cholesterol Levels HEK293 T Cells Trypsin-EDTA DMEM FBS Trypan Blue Cell Counter
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