α-Synuclein Structures Probed by 5-Fluorotryptophan Fluorescence and ^(19)F NMR Spectroscopy
Abstract
α-Synuclein, the main protein component of fibrillar deposits found in Parkinson's disease, is intrinsically disordered in vitro. Site-specific information on the protein conformation has been obtained by biosynthetic incorporation of an unnatural amino acid, 5-fluorotryptophan (5FW), into the recombinant protein. Using fluorescence and ^(19)F NMR spectroscopy, we have characterized three proteins with 5FW at positions 4, 39, and 94. Steady-state emission spectra (maxima at 353 nm; quantum yields ≈ 0.2) indicate that all three indole side chains are exposed to the aqueous medium. Virtually identical single-exponential excited-state decays (τ ≈ 3.4 ns) were observed in all three cases. Single ^(19)F NMR resonances were measured for W4, W39, and W94 at −49.0 ± 0.1 ppm. Our analysis of the spectroscopic data suggests that the protein conformations are very similar in the regions near the three sites.
Additional Information
© 2006 American Chemical Society. Received 3 January 2006. Published online 16 March 2006. Published in print 1 April 2006. This work was supported by the Arnold and Mabel Beckman Foundation (Beckman Senior Research Fellowship to J.C.L.) and the Ellison Medical Foundation (Senior Scholar Award in Aging to H.B.G.).Additional details
- Eprint ID
- 78041
- Resolver ID
- CaltechAUTHORS:20170609-072919854
- Arnold and Mabel Beckman Foundation
- Ellison Medical Foundation
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2017-06-09Created from EPrint's datestamp field
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2021-11-15Created from EPrint's last_modified field