title: Photoaffinity labeling on magnetic microspheres (PALMm) methodology for topographic mapping: preparation of PALMm reagents and demonstration of biochemical relevance.
abstract: Photoaffinity labeling (PAL) is a technique widely used for identifying the binding-site within proteins. Although the classic method is both versatile and powerful, it suffers significant disadvantages, such as the need to radiolabel the PAL ligand, and the need to conduct highly complicated separations of both the labeled protein and the labeled peptides derived from it. Here, we propose a novel and universal methodology--Photo-Affinity Labeling on Magnetic microspheres (PALMm) designed to simplify and shorten the PAL protocol. In this context, we describe the preparation of PALMm reagents and the evaluation of their biochemical relevance regarding two ATP-binding enzymes: hexokinase and apyrase.
authors.affiliation: Department of Chemistry, Gonda-Goldschmied Medical Research Center, Bar-Ilan University, Ramat-Gan 52900, Israel.
authors.forename: Efrat
authors.initials: E
authors.lastname: Halbfinger
authors.forename: Karine
authors.initials: K
authors.lastname: Gorochesky
authors.forename: Sébastien A
authors.initials: SA
authors.lastname: Lévesque
authors.forename: Adrien R
authors.initials: AR
authors.lastname: Beaudoin
authors.forename: Larisa
authors.initials: L
authors.lastname: Sheihet
authors.forename: Shlomo
authors.initials: S
authors.lastname: Margel
authors.forename: Bilha
authors.initials: B
authors.lastname: Fischer
mesh_terms: D001081:Apyrase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D015195:Drug Design; D006593:Hexokinase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D008280:Magnetics; D008863:Microspheres; D009711:Nucleotides / Q000737:chemistry / Q000378:metabolism; D019920:Photoaffinity Labels; D013056:Spectrophotometry, Ultraviolet
chemical_list: D009711:Nucleotides; D019920:Photoaffinity Labels; D006593:Hexokinase; D001081:Apyrase