Difficulties associated with the structural analysis of proteins susceptible to form aggregates: the case of tau protein as a biomarker of Alzheimer's disease
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00023752%3A_____%2F16%3A43914828" target="_blank" >RIV/00023752:_____/16:43914828 - isvavai.cz</a>
Alternative codes found
RIV/00216275:25310/16:39902239
Result on the web
<a href="http://onlinelibrary.wiley.com/doi/10.1002/jssc.201501045/epdf" target="_blank" >http://onlinelibrary.wiley.com/doi/10.1002/jssc.201501045/epdf</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1002/jssc.201501045" target="_blank" >10.1002/jssc.201501045</a>
Alternative languages
Result language
angličtina
Original language name
Difficulties associated with the structural analysis of proteins susceptible to form aggregates: the case of tau protein as a biomarker of Alzheimer's disease
Original language description
Mass spectrometry coupled with bioaffinity separation techniques is considered a powerful tool for studying protein interactions. This work is focused on epitope analysis of tau protein, which contains two VQIXXK aggregation motifs regarded as crucial elements in the formation of paired helical filaments, the main pathological characteristics of Alzheimer's disease. To identify major immunogenic structures, the epitope extraction technique utilizing protein fragmentation and magnetic microparticles functionalized with specific antibodies was applied. However, the natural adhesiveness of some newly generated peptide fragments devalued the experimental results. Beside presumed peptide fragment specific to applied monoclonal anti-tau antibodies, the epitope extraction repeatedly revealed inter alia tryptic fragment 299-HVPGGGSVQIVYKPVDLSK-317 containing the fibril-forming motif 306-VQIVYK-311. The tryptic fragment pro-aggregation and hydrophobic properties that might contribute to adsorption phenomenon were examined by Thioflavin S and reversed-phase chromatography. Several conventional approaches to reduce the non-specific fragment sorption onto the magnetic particle surface were performed, however with no effect. To avoid methodological complications, we introduced an innovative approach based on altered proteolytic digestion. Simultaneous fragmentation of tau protein by two immobilized proteases differing in the cleavage specificity (TPCK-trypsin and α-chymotrypsin) led to the disruption of motif responsible for undesirable adhesiveness and enabled us to obtain undistorted structural data.
Czech name
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Czech description
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Classification
Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
CB - Analytical chemistry, separation
OECD FORD branch
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Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2016
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Journal of Separation Science
ISSN
1615-9306
e-ISSN
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Volume of the periodical
39
Issue of the periodical within the volume
4
Country of publishing house
DE - GERMANY
Number of pages
9
Pages from-to
799-807
UT code for WoS article
000371163300023
EID of the result in the Scopus database
2-s2.0-84954290607