Research output: Contribution to journal › Article › peer-review
Estimation of amyloid aggregate sizes with semi-denaturing detergent agarose gel electrophoresis and its limitations. / Дроздова, Полина; Барбитов, Юрий Александрович; Белоусов, Михаил Владимирович; Скитченко, Ростислав; Рогоза, Татьяна Михайловна; Leclercq, Jeremy; Каява, Андрей Вилхович; Матвеенко, Андрей Георгиевич; Журавлева, Галина Анатольевна; Бондарев, Станислав Александрович.
In: Prion, Vol. 14, No. 1, 01.01.2020, p. 118-128.Research output: Contribution to journal › Article › peer-review
}
TY - JOUR
T1 - Estimation of amyloid aggregate sizes with semi-denaturing detergent agarose gel electrophoresis and its limitations
AU - Дроздова, Полина
AU - Барбитов, Юрий Александрович
AU - Белоусов, Михаил Владимирович
AU - Скитченко, Ростислав
AU - Рогоза, Татьяна Михайловна
AU - Leclercq, Jeremy
AU - Каява, Андрей Вилхович
AU - Матвеенко, Андрей Георгиевич
AU - Журавлева, Галина Анатольевна
AU - Бондарев, Станислав Александрович
N1 - Publisher Copyright: © 2020, © 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
PY - 2020/1/1
Y1 - 2020/1/1
N2 - Semi-denaturing detergent agarose gel electrophoresis (SDD-AGE) was proposed by Vitaly V. Kushnirov in the Michael D. Ter-Avanesyan’s laboratory as a method to compare sizes of amyloid aggregates. Currently, this method is widely used for amyloid investigation, but mostly as a qualitative approach. In this work, we assessed the possibilities and limitations of the quantitative analysis of amyloid aggregate size distribution using SDD-AGE results. For this purpose, we used aggregates of two well-characterized yeast amyloid-forming proteins, Sup35 and Rnq1, and developed a protocol to standardize image analysis and process the result. A detailed investigation of factors that may affect the results of SDD-AGE revealed that both the cell lysis method and electrophoresis conditions can substantially affect the estimation of aggregate size. Despite this, quantitative analysis of SDD-AGE results is possible when one needs to estimate and compare the size of aggregates on the same gel, or even in different experiments, if the experimental conditions are tightly controlled and additional standards are used.
AB - Semi-denaturing detergent agarose gel electrophoresis (SDD-AGE) was proposed by Vitaly V. Kushnirov in the Michael D. Ter-Avanesyan’s laboratory as a method to compare sizes of amyloid aggregates. Currently, this method is widely used for amyloid investigation, but mostly as a qualitative approach. In this work, we assessed the possibilities and limitations of the quantitative analysis of amyloid aggregate size distribution using SDD-AGE results. For this purpose, we used aggregates of two well-characterized yeast amyloid-forming proteins, Sup35 and Rnq1, and developed a protocol to standardize image analysis and process the result. A detailed investigation of factors that may affect the results of SDD-AGE revealed that both the cell lysis method and electrophoresis conditions can substantially affect the estimation of aggregate size. Despite this, quantitative analysis of SDD-AGE results is possible when one needs to estimate and compare the size of aggregates on the same gel, or even in different experiments, if the experimental conditions are tightly controlled and additional standards are used.
KW - Amyloids
KW - Rnq1
KW - SDD-AGE
KW - Shiny application
KW - Sup35
KW - HSP104
KW - APPEARANCE
KW - DETERMINANT
KW - SUP35
KW - PRION
KW - TRANSLATION
KW - SACCHAROMYCES-CEREVISIAE
KW - YEAST
KW - TERMINATION
KW - EXPRESSION
UR - http://www.scopus.com/inward/record.url?scp=85083645947&partnerID=8YFLogxK
UR - https://www.mendeley.com/catalogue/d9a72587-45ef-388e-9775-61b2fa03d0a8/
U2 - 10.1080/19336896.2020.1751574
DO - 10.1080/19336896.2020.1751574
M3 - Article
C2 - 32306832
VL - 14
SP - 118
EP - 128
JO - Prion
JF - Prion
SN - 1933-6896
IS - 1
ER -
ID: 52950895