Photochemical Fishing Approaches for Identifying Target Proteins and Elucidating the Structure of a Ligand-binding Region Using Carbene-generating Photoreactive Probes

Photoaffinity labeling enables the direct probing of a target protein through a covalent bond between a ligand and its binding protein, and even a complex formed by weak interactions can be isolated by the method. The photochemical fishing approach accelerates the throughput, isolating crosslinked c...

Full description

Saved in:
Bibliographic Details
Published in:Analytical Sciences 2006, Vol.22(2), pp.209-218
Main Authors: SADAKANE, Yutaka, HATANAKA, Yasumaru
Format: Article
Language:English
Subjects:
Azides - chemistry
Biotinylation
Carbohydrates - chemistry
DNA - chemistry
Hydrocarbons - chemistry
Ligands
Light
Methane - analogs & derivatives
Methane - chemistry
Microscopy, Fluorescence - instrumentation
Microscopy, Fluorescence - methods
Models, Biological
Models, Chemical
Peptides - chemistry
Photoaffinity Labels - chemistry
Photochemistry - methods
Proteins - chemistry
RNA, Transfer - chemistry
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Photoaffinity labeling enables the direct probing of a target protein through a covalent bond between a ligand and its binding protein, and even a complex formed by weak interactions can be isolated by the method. The photochemical fishing approach accelerates the throughput, isolating crosslinked complexes and analyzing the structure of the ligand binding site within the protein. We used carbene-generating phenyldiazirine for this approach because practical examinations had shown that the phenyldiazirine functioned as the powerful barb on the hook. Improving the synthetic pathways of the photoprobes and using chemoselective-integrated photoreactive units makes possible the easy and rapid preparation of carbene-generating photoreactive probes including the derivatives in peptides, proteins, DNAs, and carbohydrates. This review also shows several recent impacts of photoaffinity labeling, including the in vivo preparation of photoreactive proteins in living cells.
ISSN:0910-6340
1348-2246
DOI:10.2116/analsci.22.209