Biotinylated anisomycin: a comparison of classical and "click" chemistry approaches.
Two approaches to the synthesis of biotinylated derivatives of the stress-activated protein kinase (SAPK) pathway activator anisomycin have been investigated. Attachment of the biotin moiety to the central core was achieved either through the use of a classical displacement reaction on alpha-halo carbonyl derivatives of biotin or through a copper(I)-catalyzed 1,3-dipolar Huisgen cycloaddition ("click") coupling of biotinylated azides to propargyl-marked analogues of anisomycin. In each case, the resultant N-linked molecular probes were found to be active in SAPK pathway immunoblot assays, while their O-linked counterparts were inactive. However, in sharp contrast to the classical coupling approach which results in low coupling yields, the aqueous "click" coupling process was found to deliver high yields of biotinylated probes, making it the conjugation method of choice. A survey of the available methods for the addition of a propargyl marker onto a range of chemical functionalities strongly suggests that this copper(I)-catalyzed 1,3-dipolar Huisgen cycloaddition approach to biotinylation may be generally applied.
Inverarity
Iain A
IA
The School of Chemistry, University of Edinburgh, Kings Buildings, West Mains Road, Edinburgh, EH9 3JJ, United Kingdom.
Viguier
Romain F H
RF
Cohen
Philip
P
Hulme
Alison N
AN
D000841:Anisomycin / Q000031:analogs & derivatives / Q000138:chemical synthesis* / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000494:pharmacology; D001386:Azides / Q000737:chemistry; D019921:Biotinylation; D002384:Catalysis; D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D003300:Copper / Q000737:chemistry; D015151:Immunoblotting / Q000379:methods; D015394:Molecular Structure; D018384:Oxidative Stress; D010293:Pargyline / Q000737:chemistry; D011494:Protein Kinases / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D001386:Azides; D000841:Anisomycin; D003300:Copper; D010293:Pargyline; D011494:Protein Kinases