Photoaffinity Heterobifunctional Cross-Linking Reagents Based on Azide-Substituted Salicylates

被引:3
作者
Imai, Nobuyuki [2 ]
Dwyer, Lori D. [1 ]
Kometani, Tadashi [2 ]
Ji, Tae [3 ]
Vanaman, Thomas C. [1 ]
Watt, David S. [2 ]
机构
[1] Univ Kentucky, Dept Biochem, Lexington, KY 40506 USA
[2] Univ Kentucky, Dept Chem, Lexington, KY 40506 USA
[3] Univ Wyoming, Dept Biochem, Laramie, WY 82071 USA
基金
美国国家科学基金会;
关键词
D O I
10.1021/bc00002a009
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:144 / 148
页数:5
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