Photoaffinity labeling on magnetic microspheres (PALMm) methodology for topographic mapping: Preparation of PALMm reagents and demonstration of biochemical relevance

Efrat Halbfinger, Karine Gorochesky, Sébastien A. Lévesque, Adrien R. Beaudoin, Larisa Sheihet, Shlomo Margel, Bilha Fischer

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

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.

Original languageEnglish
Pages (from-to)2821-2832
Number of pages12
JournalOrganic and Biomolecular Chemistry
Volume1
Issue number16
DOIs
StatePublished - 21 Aug 2003

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