Impact of Deuteration on the Assembly Kinetics of Transthyretin Monitored by Native Mass Spectrometry and Implications for Amyloidoses.
It is well established that the formation of transthyretin (TTR) amyloid fibrils is linked to the destabilization and dissociation of its tetrameric structure into insoluble aggregates. Isotope labeling is used for the study of TTR by NMR, neutron diffraction, and mass spectrometry (MS). Here MS, thioflavin T fluorescence, and crystallographic data demonstrate that while the X-ray structures of unlabeled and deuterium-labeled TTR are essentially identical, subunit exchange kinetics and amyloid formation are accelerated for the deuterated protein. However, a slower subunit exchange is noted in deuterated solvent, reflecting the poorer solubility of non-polar protein side chains in such an environment. These observations are important for the interpretation of kinetic studies involving deuteration. The destabilizing effects of TTR deuteration are rather similar in character to those observed for aggressive mutations of TTR such as L55P (associated with familial amyloid polyneuropathy).
Life Sciences group, ILL, 71 avenue des Martyrs, 38042, Grenoble, France.|Faculty of Natural Sciences, Keele University, Staffordshire, ST5 5BG, UK.
Ai Woon
0000-0002-2137-041X
AW
Yee
Life Sciences group, ILL, 71 avenue des Martyrs, 38042, Grenoble, France.|Faculty of Natural Sciences, Keele University, Staffordshire, ST5 5BG, UK.
Martine
M
Moulin
Life Sciences group, ILL, 71 avenue des Martyrs, 38042, Grenoble, France.
Nina
N
Breteau
Life Sciences group, ILL, 71 avenue des Martyrs, 38042, Grenoble, France.
Michael
M
Haertlein
Faculty of Natural Sciences, Keele University, Staffordshire, ST5 5BG, UK.|ESRF, 71 avenue des Martyrs, 38042, Grenoble, France.
Edward P
EP
Mitchell
Laboratory of Protein Crystallography, Drug Discovery Group, Wolfson Institute for Biomedical Research, UCL, London, WC1E 6BT, UK.
Jonathan B
JB
Cooper
Univ. Grenoble Alpes, IBS, 38044, Grenoble, France. elisabetta.boeri-erba@ibs.fr.|CNRS, IBS, 38044, Grenoble, France. elisabetta.boeri-erba@ibs.fr.|CEA, IBS, 38044, Grenoble, France. elisabetta.boeri-erba@ibs.fr.
Elisabetta
0000-0002-5910-4707
E
Boeri Erba
Life Sciences group, ILL, 71 avenue des Martyrs, 38042, Grenoble, France. tforsyth@ill.fr.|Faculty of Natural Sciences, Keele University, Staffordshire, ST5 5BG, UK. tforsyth@ill.fr.
V Trevor
VT
Forsyth
D000686:Amyloidosis / Q000378:metabolism*; D052160:Benzothiazoles; D018360:Crystallography, X-Ray; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D007553:Isotope Labeling; D007700:Kinetics; D013058:Mass Spectrometry; D008958:Models, Molecular; D011228:Prealbumin / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D013844:Thiazoles / Q000737:chemistry
D052160:Benzothiazoles; D005456:Fluorescent Dyes; D011228:Prealbumin; D013844:Thiazoles; C009462:thioflavin T
amyloid proteins; deuteration; isotope effects; native mass spectrometry; transthyretin