The smallest resonance energy transfer acceptor for tryptophan.
The utility of diazirine ligands as acceptors in resonance energy transfer (RET) distance measurements with tryptophan or tryptophan analogues as donor is reported. The principle is demonstrated for a diazirine derivative of d-mannitol, 2-azi-2-deoxy-d-arabino-hexitol, and single-tryptophan-containing mutants of the mannitol transporter, EIImtl, from E. coli. The Förster distance is 10 A for the tryptophan-diazirine donor-acceptor couple, allowing the measurement of distances up to 17 A. The versatility of tryptophan as an intrinsic spectroscopic probe in protein chemistry and the small size of the diazirine group makes this a very attractive donor-acceptor couple for accurate RET distance information in protein chemistry.
Broos
Jaap
J
Department of Biochemistry and Groningen Biomolecular Science and Biotechnology Institute (GBB), University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands. J.Broos@chem.rug.nl
Pas
Hendri H
HH
Robillard
George T
GT
D001089:Arabinose / Q000031:analogs & derivatives / Q000737:chemistry*; D001389:Azirines / Q000737:chemistry*; D001665:Binding Sites; D004926:Escherichia coli / Q000737:chemistry; D029968:Escherichia coli Proteins; D008353:Mannitol / Q000031:analogs & derivatives / Q000378:metabolism; D009004:Monosaccharide Transport Proteins; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000737:chemistry*
C462102:2-azi-2-deoxy-arabino-hexitol; D001389:Azirines; D029968:Escherichia coli Proteins; D009004:Monosaccharide Transport Proteins; D008353:Mannitol; D014364:Tryptophan; D001089:Arabinose; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C022358:mannitol PTS permease, E coli