A live cell, image-based approach to understanding the enzymology and pharmacology of 2-bromopalmitate and palmitoylation.

摘要: Abstract The addition of a lipid moiety to protein increases its hydrophobicity and subsequently attraction lipophilic environments like membranes. Indeed most lipid‐modified proteins are localized membranes where they associate with multiprotein signaling complexes. Acylation prenylation the two common categories lipidation. enzymology pharmacology well understood but relatively very little is known about palmitoylation, form acylation. One distinguishing characteristic palmitoylation that it dynamic modification. To understand more how regulated, we fused substrates fluorescent reported their subcellular distribution trafficking. We used automated high‐throughput fluorescence microscopy specialized computer algorithm image measure fraction reporter on plasma membrane versus cytoplasm. Using this system determined residence half‐life palmitate dipalmitoyl substrate peptide from GAP43 as EC 50 for 2‐bromopalmitate, inhibitor palmitoylation.