Letter abstract
Nature Biotechnology 25, 1483 - 1487 (2007)
Published online: 2 December 2007 | doi:10.1038/nbt1355
Redirecting lipoic acid ligase for cell surface protein labeling with small-molecule probes
Marta Fernández-Suárez1, Hemanta Baruah1, Laura Martínez-Hernández1, Kathleen T Xie1, Jeremy M Baskin2, Carolyn R Bertozzi2,3,4,5 & Alice Y Ting1
Live cell imaging is a powerful method to study protein dynamics at the cell surface, but conventional imaging probes are bulky, or interfere with protein function1, 2, or dissociate from proteins after internalization3, 4. Here, we report technology for covalent, specific tagging of cellular proteins with chemical probes. Through rational design, we redirected a microbial lipoic acid ligase (LplA)5 to specifically attach an alkyl azide onto an engineered LplA acceptor peptide (LAP). The alkyl azide was then selectively derivatized with cyclo-octyne6 conjugates to various probes. We labeled LAP fusion proteins expressed in living mammalian cells with Cy3, Alexa Fluor 568 and biotin. We also combined LplA labeling with our previous biotin ligase labeling7, 8, to simultaneously image the dynamics of two different receptors, coexpressed in the same cell. Our methodology should provide general access to biochemical and imaging studies of cell surface proteins, using small fluorophores introduced via a short peptide tag.
- Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, Massachusetts 02139, USA.
- Department of Chemistry, University of California, Berkeley, California 94720, USA.
- Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA.
- Howard Hughes Medical Institute, University of California, Berkeley, California 94720, USA.
- The Molecular Foundry, Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA.
Correspondence to: Alice Y Ting1 e-mail: ating@mit.edu
MORE ARTICLES LIKE THIS
These links to content published by NPG are automatically generated.
RESEARCH
Hypothetical biotechnology mega-mergersNature Biotechnology Strategic Planning (01 May 1999)
Hypothetical biotechnology mega-mergersNature Biotechnology Strategic Planning (01 May 1999)
See all 9 matches for Research
