Biochemistry (Moscow)

, Volume 72, Issue 1, pp 1–20

Photoaffinity labeling and its application in structural biology

Authors

  • E. L. Vodovozova
    • Shemyakin and Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of Sciences
Review

DOI: 10.1134/S0006297907010014

Cite this article as:
Vodovozova, E.L. Biochemistry (Moscow) (2007) 72: 1. doi:10.1134/S0006297907010014

Abstract

This review contains a brief consideration of some theoretical aspects of photoaffinity (photoreactive) labeling (PAL), and the most widely used photoreactive groups, such as arylazide, benzophenone, and 3-(trifluoromethyl)-3-phenyldiazirine, are characterized in comparison. Experimental methodology is described, including modern approaches of mass spectrometry for analysis of cross-linking products between the photoreactive probes and biomolecules. Examples of PAL application in diverse fields of structural biology during the last five-ten years are presented. Potential drug targets, transport processes, stereochemistry of interaction of G-protein-coupled receptors with ligands, as well as structural changes in nicotinic acetylcholine receptor are considered. Applications of photoaffinity ganglioside and phospholipid probes for studying biological membranes and of nucleotide probes in investigations of replicative and transcriptional complexes, as well as photoaffinity glycoconjugates for detecting carbohydrate-binding proteins are covered. In combination with modern techniques of instrumental analysis and computer-aided modeling, PAL remains the most important approach in studies on the organization of biological systems.

Key words

photoaffinity labelingligand-binding sitesphotoreactive probesgangliosidesnucleotidesglycoconjugates

Abbreviations

Bp

benzoylphenyl

Bpa

p-benzoylphenylalanine

CRF

corticotropin-releasing factor

DAT

dopamine protein transporter

Dcp

diazocyclopentadien-2-ylcarbonyl

ESI

electrospray ionization

GPCR

G-protein-coupled receptor

GSph

gangliosides

ILBP

ileal lipid-binding protein

LC/MS

liquid chromatography/mass spectrometry

MALDI

matrix assisted laser desorption ionization

MDR

multiple drug resistance

MS

mass spectrometry

MS/MS

tandem mass spectrometry

nAChR

nicotinic acetylcholine receptor

Nab

2-nitro-5-azidobenzoyl

Nap

2-nitro-4-azidophenyl

PAL

photoaffinity labeling

PC

phosphatidylcholine

SE

squalene epoxidase

TD

transmembrane domain

Tfd

3-(trifluoromethyl)-3-phenyldiazirine

Copyright information

© Pleiades Publishing, Ltd. 2007