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The smallest resonance energy transfer acceptor for tryptophan
Jaap Broos1, Hendri H Pas, George T Robillard
1Department 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
Journal of the American Chemical Society
|June 13, 2002
Summary
This study introduces diazirine ligands for resonance energy transfer (RET) measurements, using tryptophan as a donor. This novel approach provides accurate distance information in protein chemistry.
Area of Science:
- Biophysical Chemistry
- Protein Science
- Spectroscopy
Background:
- Resonance energy transfer (RET) is a powerful technique for measuring distances within biological molecules.
- Tryptophan is a versatile intrinsic fluorophore used as a donor in RET studies.
- Diazirine ligands offer potential as efficient energy acceptors.
Purpose of the Study:
- To evaluate the utility of diazirine ligands as acceptors for RET distance measurements.
- To demonstrate the application of tryptophan-diazirine donor-acceptor pairs in protein science.
- To establish a method for obtaining accurate distance information in complex biological systems.
Main Methods:
- Utilized diazirine ligands as resonance energy transfer (RET) acceptors.
- Employed tryptophan and its analogues as spectroscopic donors.
- Investigated a diazirine derivative of d-mannitol and single-tryptophan mutants of the mannitol transporter (EIImtl) from E. coli.
- Applied Förster resonance energy transfer (FRET) principles for distance determination.
Main Results:
- Established the tryptophan-diazirine donor-acceptor couple for RET measurements.
- Determined a Förster distance of 10 Å for this pair.
- Demonstrated the capability to measure distances up to 17 Å.
- Validated the method using a model compound and a protein system.
Conclusions:
- Diazirine ligands are effective acceptors for RET distance measurements with tryptophan donors.
- The small size of diazirine and tryptophan's spectroscopic properties make them an attractive pair for protein chemistry.
- This method enables accurate RET distance information crucial for understanding protein structure and function.
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