Raman spectral imaging of 13 C 2 H 15 N-labeled α-synuclein amyloid fibrils in cells.

Autor: Watson MD; Laboratory of Protein Conformation and Dynamics, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, United States of America., Flynn JD; Laboratory of Protein Conformation and Dynamics, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, United States of America., Lee JC; Laboratory of Protein Conformation and Dynamics, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, United States of America. Electronic address: leej4@nhlbi.nih.gov.
Jazyk: angličtina
Zdroj: Biophysical chemistry [Biophys Chem] 2021 Feb; Vol. 269, pp. 106528. Date of Electronic Publication: 2020 Dec 14.
DOI: 10.1016/j.bpc.2020.106528
Abstrakt: Parkinson's disease is characterized by the intracellular accumulation of α-synuclein (α-syn) amyloid fibrils, which are insoluble, β-sheet-rich protein aggregates. Raman spectroscopy is a powerful technique that reports on intrinsic molecular vibrations such as the coupled vibrational modes of the polypeptide backbone, yielding secondary structural information. However, in order to apply this method in cells, spectroscopically unique frequencies are necessary to resolve proteins of interest from the cellular proteome. Here, we report the use of 13 C 2 H 15 N-labeled α-syn to study the localization of preformed fibrils fed to cells. Isotopic labeling shifts the amide-I ( 13 CO) band away from endogenous 12 CO vibrations, permitting secondary structural analysis of internalized α-syn fibrils. Similarly, 13 C 2 H stretches move to lower energies in the "cellular quiet" region, where there is negligible biological spectral interference. This combination of well-resolved, distinct vibrations allows Raman spectral imaging of α-syn fibrils across a cell, which provides conformational information with spatial context.
(Published by Elsevier B.V.)
Databáze: MEDLINE
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