Реагент Денни-Джеффа

Description

The Denny-Jaffe reagent is a heterobifunctional cleavable photoactivated cross-linking reagent used in biochemistry for photoaffinity labeling in the study of interactions of proteins with each other and with the cell surface. Chemically it is N-(4-(p-azido-m-iodophenylazo)benzoyl)-3-aminopropyl-N'-hydroxysulfosuccinimide ester, first described by Denny and Blobel. The molecule contains a photoreactive aryl azide group and an azo bond, sensitive to reductive cleavage, as well as iodine, which, after photolysis and cleavage of the azo bond, is transferred directly to the target molecule cross-linked with the reagent.

Basic Attributes

• CAS No: 109545-43-9
• Chemical formula: C20H16IN7O5
• Molecular weight: 561.28941
• Exact weight: 561.026
• PSA Specifications: 132
• XLogP3: 3.5

Operating principle

The reagent contains an N-hydroxysuccinimide ester that reacts with the primary amino groups (eg, lysine residues) of the target protein and covalently attaches to it the rest of the molecule, including the photoactivatable aryl azide group. When irradiated with ultraviolet light, the azide group is converted into a highly reactive nitrene, which is capable of binding to nearby molecules - neighboring proteins or sections of the cell membrane - thereby fixing the spatial arrangement of interacting partners. A key feature of the reagent is the cleavable azo bond: once a cross-link has formed, it can be selectively broken by a reducing agent, releasing the labeled molecule and transferring a radioactive or other label (in this case, an iodine atom) directly to the interaction partner, facilitating subsequent identification.

Scope of application

• photoaffinity labeling for mapping protein-protein interactions
• study of cell surface receptors and their ligands
• studies of the spatial organization of membrane protein complexes
• transfer of a radioactive label (iodine) to an interaction partner for subsequent detection

Importance of Photoaffine Labeling Method

Photoaffinity labeling is a biochemical technique that uses cross-linkers with photoreactive groups (most often aryl azide) to obtain three-dimensional information about the binding surfaces between proteins or between a protein and a ligand. Cleavable reagents like the Denny-Jeff reagent are particularly valuable because they allow, after formation of a covalent cross-link, the initially bound molecules to be separated and the interaction partner to be unambiguously identified—this greatly simplifies the interpretation of results compared to non-cleavable cross-linkers.

Storage and transportation conditions

Store in a place protected from light (the aryl azide group is photosensitive) at a low temperature, in an airtight container. Working with the reagent requires darkened conditions until controlled photoirradiation in the experiment.