Microfluidic Mobility Shift Assay for Real-Time Analysis of Peptide N-Palmitoylation.

ABSTRACT: The Hedgehog pathway is a key developmental signaling pathway but is also implicated in many types of cancer. The extracellular signaling protein Sonic hedgehog (Shh) requires dual lipidation for functional signaling, whereby N-terminal palmitoylation is performed by the enzyme Hedgehog acyltransferase (Hhat). Hhat is an attractive target for small-molecule inhibition to arrest Hedgehog signaling, and methods for assaying Hhat activity are central to understanding its function. However, all existing assays to quantify lipidation of peptides suffer limitations, such as safety hazards, high costs, extensive manual handling, restriction to stopped-assay measurements, or indirect assessment of lipidation. To address these limitations, we developed a microfluidic mobility shift assay (MSA) to a

SUBMITTER: Lanyon-Hogg T 

PROVIDER: S-EPMC5453399 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature