Probing protein electrostatics with a synthetic fluorescent amino acid

被引:352
作者
Cohen, BE [1 ]
McAnaney, TB
Park, ES
Jan, YN
Boxer, SG
Jan, LY
机构
[1] Univ Calif San Francisco, Howard Hughes Med Inst, San Francisco, CA 94143 USA
[2] Univ Calif San Francisco, Dept Physiol, San Francisco, CA 94143 USA
[3] Univ Calif San Francisco, Dept Biochem, San Francisco, CA 94143 USA
[4] Stanford Univ, Dept Chem, Stanford, CA 94305 USA
关键词
D O I
10.1126/science.1069346
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Electrostatics affect virtually all aspects of protein structure and activity and are particularly important in proteins whose primary function is to stabilize charge. Here we introduce a fluorescent amino acid, Aladan, which can probe the electrostatic character of a protein at multiple sites. Aladan is exceptionally sensitive to the polarity of its surroundings and can be incorporated site-selectively at buried and exposed sites, in both soluble and membrane proteins. Steady-state and time-resolved fluorescence measurements of Aladan residues at different buried and exposed sites in the B1 domain of protein G suggest that its interior is polar and heterogeneous.
引用
收藏
页码:1700 / 1703
页数:4
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