Photoaffinity heterobifunctional cross-linking reagents based on azide-substituted salicylates.
Photoaffinity heterobifunctional cross-linking reagents are described that incorporate a 4-azidosalicylate group at one terminus and an N-hydroxysuccinimidyl ester at the other terminus with a "linking arm" of variable length separating the photoactive and electrophilic termini. Exposure of calmodulin (CaM) to succinimidyl N-[2-[(4-azidosalicyloyl)oxy]ethyl]suberamate (4C) led to monoadducts at Lys-21, Lys-75, and Lys-94. Separation of the monoadducts from CaM and polyadducts, radioiodination, and photolysis in the presence of human erythrocyte plasma membrane Ca2+, Mg2(+)-ATPase led to calcium-dependent cross-linking with 8% cross-linking efficiency.
Imai
N
N
Department of Chemistry, University of Kentucky, Lexington 40506.
Dwyer
L D
LD
Kometani
T
T
Ji
T
T
Vanaman
T C
TC
Watt
D S
DS
D000345:Affinity Labels / Q000138:chemical synthesis* / Q000737:chemistry; D001386:Azides; D017301:Ca(2+) Mg(2+)-ATPase / Q000097:blood; D000252:Calcium-Transporting ATPases / Q000097:blood; D002147:Calmodulin / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000138:chemical synthesis* / Q000737:chemistry; D004910:Erythrocyte Membrane / Q000201:enzymology; D006801:Humans; D007202:Indicators and Reagents; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D011485:Protein Binding; D012459:Salicylates; D013055:Spectrophotometry, Infrared
D000345:Affinity Labels; D001386:Azides; D002147:Calmodulin; D003432:Cross-Linking Reagents; D007202:Indicators and Reagents; D012459:Salicylates; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases