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Science 31 May 2002:
Vol. 296. no. 5573, pp. 1700 - 1703
DOI: 10.1126/science.1069346
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Reports

Probing Protein Electrostatics with a Synthetic Fluorescent Amino Acid

Bruce E. Cohen,1dagger Tim B. McAnaney,2* Eun Sun Park,2* Yuh Nung Jan,1 Steven G. Boxer,2 Lily Yeh Jan1

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.

1 Howard Hughes Medical Institute and Departments of Physiology and Biochemistry, University of California San Francisco, San Francisco, CA 94143, USA.
2 Department of Chemistry, Stanford University, Stanford, CA 94305, USA.
*   These authors contributed equally to this work.

dagger    To whom correspondence should be addressed. Email: bcohen{at}itsa.ucsf.edu

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