Reversible site-selective labeling of membrane proteins in live cells - PubMed (original) (raw)
doi: 10.1038/nbt954. Epub 2004 Mar 21.
- PMID: 15034592
- DOI: 10.1038/nbt954
Reversible site-selective labeling of membrane proteins in live cells
Emmanuel G Guignet et al. Nat Biotechnol. 2004 Apr.
Abstract
Chemical and biological labeling is fundamental for the elucidation of the function of proteins within biochemical cellular networks. In particular, fluorescent probes allow detection of molecular interactions, mobility and conformational changes of proteins in live cells with high temporal and spatial resolution. We present a generic method to label proteins in vivo selectively, rapidly (seconds) and reversibly, with small molecular probes that can have a wide variety of properties. These probes comprise a chromophore and a metal-ion-chelating nitrilotriacetate (NTA) moiety, which binds reversibly and specifically to engineered oligohistidine sequences in proteins of interest. We demonstrate the feasibility of the approach by binding NTA-chromophore conjugates to a representative ligand-gated ion channel and G protein-coupled receptor, each containing a polyhistidine sequence. We investigated the ionotropic 5HT(3) serotonin receptor by fluorescence measurements to characterize in vivo the probe-receptor interactions, yielding information on structure and plasma membrane distribution of the receptor.
Comment in
- Finding needles in haystacks.
Javitch JA. Javitch JA. Nat Biotechnol. 2004 Apr;22(4):394-6. doi: 10.1038/nbt0404-394. Nat Biotechnol. 2004. PMID: 15060551 No abstract available.
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