Correctors of protein trafficking defects identified by a novel high-throughput screening assay.
High-throughput small-molecule screens hold great promise for identifying compounds with potential therapeutic value in the treatment of protein-trafficking diseases such as cystic fibrosis (CF) and nephrogenic diabetes insipidus (NDI). The approach usually involves expressing the mutant form of the gene in cells and assaying function in a multiwell format when cells are exposed to libraries of compounds. Although such functional assays are useful, they do not directly test the ability of a compound to correct defective trafficking of the protein. To address this we have developed a novel corrector-screening assay for CF, in which the appearance of the mutant protein at the cell surface is measured. We used this assay to screen a library of 2000 compounds and have isolated several classes of trafficking correctors that had not previously been identified. This novel screening approach to protein-trafficking diseases is robust and general, and could enable the selection of molecules that could be translated rapidly to a clinical setting.
Carlile
Graeme W
GW
Department of Biochemistry, McGill University, McIntyre Medical Building, 3655 Promenade Sir William Osler, Montréal, QC, H3G 1Y6, Canada.
Robert
Renaud
R
Zhang
Donglei
D
Teske
Katrina A
KA
Luo
Yishan
Y
Hanrahan
John W
JW
Thomas
David Y
DY
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002463:Cell Membrane Permeability; D006224:Cricetinae; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000737:chemistry; D004353:Drug Evaluation, Preclinical; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006388:Hemagglutinins / Q000737:chemistry; D007454:Iodides / Q000737:chemistry; D010879:Piperazines / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D011687:Purines / Q000737:chemistry; D000068677:Sildenafil Citrate; D013050:Spectrometry, Fluorescence; D013450:Sulfones / Q000737:chemistry; D016707:Tissue Fixation
D006388:Hemagglutinins; D007454:Iodides; D010879:Piperazines; D011506:Proteins; D011687:Purines; D013450:Sulfones; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000068677:Sildenafil Citrate