ArticleProceedings of the National Academy of Sciences of the United States of America2002
Engineering a terbium-binding site into an integral membrane protein for luminescence energy transfer.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 58 citations in OpenAlex.
- Protein and peptide engineering for chemical exchange saturation transfer imaging in the age of synthetic biology.NMR in biomedicine · 2023Review
- Photoluminescence of Homoleptic Lanthanide Complexes With Tris(benzotriazol-1-yl)borate.Journal of fluorescence · 2021Article
- Article
- Probing Polytopic Membrane Protein-Substrate Interactions by Luminescence Resonance Energy Transfer.Journal of the American Chemical Society · 2016Article
- Sensitized near-infrared luminescence from Nd(III), Yb(III) and Er(III) complexes by energy-transfer from ruthenium 1,3-Bis([1,10]phenanthroline-[5,6-d]-imidazol-2 -yl)benzene.Journal of fluorescence · 2013Article
- Binding of ReO4(-) with an engineered MoO4(2-)-binding protein: towards a new approach in radiopharmaceutical applications.Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry · 2012Article
- Structural identification of cation binding pockets in the plasma membrane proton pump.Proceedings of the National Academy of Sciences of the United States of America · 2010Article
- Single-molecule imaging of a fluorescent unnatural amino acid incorporated into nicotinic receptors.Biophysical journal · 2009Article
- Monitoring pyrene excimers in lactose permease liposomes: revealing the presence of phosphatidylglycerol in proximity to an integral membrane protein.Journal of fluorescence · 2007Article
- Change in permeant size selectivity by phosphorylation of connexin 43 gap-junctional hemichannels by PKC.Proceedings of the National Academy of Sciences of the United States of America · 2007Article
- Phosphorylation-dependent metal binding by alpha-synuclein peptide fragments.Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry · 2007Article
- Imaging single cardiac ryanodine receptor Ca2+ fluxes in lipid bilayers.Biophysical journal · 2004Article
- Lanthanide-binding helix-turn-helix peptides: solution structure of a designed metallonuclease.Proceedings of the National Academy of Sciences of the United States of America · 2003Article
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
Abstract
Luminescence resonance energy transfer with a lanthanide like Tb(3+) as donor is a useful technique for estimating intra- and intermolecular distances in macromolecules. However, the technique usually requires the use of a bulky chelator with a flexible linker attached to a Cys residue to bind Tb(3+) and, for intramolecular studies, an acceptor fluorophor attached to another Cys residue in the same protein. Here, an engineered EF- hand motif is incorporated into the central cytoplasmic loop of the lactose permease of Escherichia coli generating a high-affinity site for Tb(3+) (K(Tb)(3+) approximately 4.5 microM) or Gd(3+) (K(Gd)(3+) approximately 2.3 microM). By exciting a Trp residue in the coordination sequence, Tb(3+) bound to the EF-hand motif is sensitized specifically, and the efficiency of energy transfer to strategically placed Cys residues labeled with fluorophors is measured. In this study, we use the technique to measure distance from the EF-hand in the central cytoplasmic loop of lactose permease to positions 179 or 169 at the center or periplasmic end of helix VI, respectively. The average calculated distances of approximately 23 A (position 179) and approximately 33 A (position 169) observed with three different fluorophors as acceptors agree well with the geometry of a slightly tilted alpha-helix. The approach should be of general use for studying static and dynamic aspects of polytopic membrane protein structure and function.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.