Broos, Jaap

Pas, Hendri H.

Robillard, George T.

University of Groningen

   University of GroningenThe Smallest Resonance Energy Transfer Acceptor for TryptophanBroos, Jaap; Pas, Hendri H.; Robillard, George T.Published in:Biophysical JournalIMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite fromit. Please check the document version below.Document VersionPublisher's PDF, also known as Version of recordPublication date:2002Link to publication in University of Groningen/UMCG research databaseCitation for published version (APA):Broos, J., Pas, H. H., & Robillard, G. T. (2002). The Smallest Resonance Energy Transfer Acceptor forTryptophan. Biophysical Journal.CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of theauthor(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).Take-down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediatelyand investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons thenumber of authors shown on this cover page is limited to 10 maximum.Download date: 12-11-2019THE SMALLEST RESONANCE ENERGY TRANSFER ACCEPTOR FOR TRYPTOPHAN Jaap Broos, Erwin P.P. Vos, Hendri H. Pas, George T. Robillard; University of Groningen, Nijenborgh 4, Groningen, 9747 AG Netherlands Diazirine compounds have found widespread use as photoactivatable reagents. In this work we report on the extension of the utility of diazirine ligands in protein science. The diazirine moiety, which absorb between 330-380 nm, is used as an acceptor for tryptophan (Trp) and Trp analogs, like 5-fluoroTrp, in resonance energy transfer (RET) experiments. Distances up to 18 Χ can be determined with high accuracy. The principle is demonstrated for a diazirine derivative of mannitol, 2-azi-2-deoxy-D-arabino-hexitol (azi-mannitol) and a dozen single-Trp-containing mutants of the membrane bound mannitol transporter, EIImtl, from E. coli. The diazirine moiety is photostable under the experimental conditions. The ambiguity about the exact position of the chromophoric group relative to the ligand due to the use of a flexible linker is absent in azi-mannitol, since the chromophoric group is an integral part of the substrate. The small size of the diazirine group allows its isosteric introduction in biological ligands without a large effect on the biological activity. In conclusion, the versatility of Trp as an intrinsic spectroscopic probe of proteins and the small size of the diazirine group make this a very attractive donor-acceptor couple for accurate RET distance information in protein chemistry.   

The Smallest Resonance Energy Transfer Acceptor for Tryptophan

https://pure.rug.nl/ws/files/17486040/2002BiophysJBroos.pdf

The Smallest Resonance Energy Transfer Acceptor for Tryptophan

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