p-Nitrophenyl 3-diazopyruvate and diazopyruvamides, a new family of photoactivatable cross-linking bioprobes.
p-Nitrophenyl 3-diazopyruvate (DAPpNP) has been developed as a heterobifunctional cross-linking agent for synthesis of photoaffinity probes and photoactivatable cross-linking agents that are nucleophile specific. p-Nitrophenyl chloroglyoxylate is formed in high yield from oxalyl chloride and p-nitrophenol. Subsequent reaction with diazomethane produces DAPpNP in 50-60% overall yield. DAPpNP acylates primary and secondary amines to form 3-diazopyruvamides in high yields. 3-Diazopyruvamide derivatives have been formed from a wide variety of amines including aromatic amines, amino acids, and peptides. 3-Diazopyruvamides undergo photolysis and Wolff rearrangement at 300 nm to produce a ketene amide, which efficiently acylates nucleophilic species to form malonic acid amide derivatives. A family of photoactivatable 3-diazopyruvamide cross-linking agents was synthesized from amino acids. A cleavable, thiol-specific photoactivatable cross-linking agent was synthesized from cystamine. These reagents were caused to react with rabbit muscle aldolase to form mainly dimeric cross-linked species.
Goodfellow
V S
VS
Department of Chemistry, University of Michigan, Ann Arbor 48109.
Settineri
M
M
Lawton
R G
RG
D003432:Cross-Linking Reagents / Q000138:chemical synthesis*; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D007202:Indicators and Reagents; D009682:Magnetic Resonance Spectroscopy; D008970:Molecular Weight; D009596:Nitrophenols / Q000138:chemical synthesis* / Q000494:pharmacology; D010782:Photolysis; D011773:Pyruvates / Q000138:chemical synthesis* / Q000494:pharmacology; D013053:Spectrophotometry; D013329:Structure-Activity Relationship; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D007202:Indicators and Reagents; D009596:Nitrophenols; D011773:Pyruvates; C061076:4-nitrophenyl 3-diazopyruvate; D005634:Fructose-Bisphosphate Aldolase