A method of predicting the in vitro fibril formation propensity of Abeta40 mutants based on their inclusion body levels in E. coli

Kalyani Sanagavarapu, Elisabeth Nüske, Irem Nasir, Georg Meisl, Jasper N. Immink, Pietro Sormanni, Michele Vendruscolo, Tuomas P.J. Knowles, Anders Malmendal, Celia Cabaleiro-Lago, Sara Linse

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Overexpression of recombinant proteins in bacteria may lead to their aggregation and deposition in inclusion bodies. Since the conformational properties of proteins in inclusion bodies exhibit many of the characteristics typical of amyloid fibrils. Based on these findings, we hypothesize that the rate at which proteins form amyloid fibrils may be predicted from their propensity to form inclusion bodies. To establish a method based on this concept, we first measured by SDS-PAGE and confocal microscopy the level of inclusion bodies in E. coli cells overexpressing the 40-residue amyloid-beta peptide, Aβ40, wild-type and 24 charge mutants. We then compared these results with a number of existing computational aggregation propensity predictors as well as the rates of aggregation measured in vitro for selected mutants. Our results show a strong correlation between the level of inclusion body formation and aggregation propensity, thus demonstrating the power of this approach and its value in identifying factors modulating aggregation kinetics.
OriginalsprogEngelsk
Artikelnummer3680
TidsskriftScientific Reports
Vol/bind9
Udgave nummer1
ISSN2045-2322
DOI
StatusUdgivet - 2019
Udgivet eksterntJa

Citer dette

Sanagavarapu, Kalyani ; Nüske, Elisabeth ; Nasir, Irem ; Meisl, Georg ; Immink, Jasper N. ; Sormanni, Pietro ; Vendruscolo, Michele ; Knowles, Tuomas P.J. ; Malmendal, Anders ; Cabaleiro-Lago, Celia ; Linse, Sara. / A method of predicting the in vitro fibril formation propensity of Abeta40 mutants based on their inclusion body levels in E. coli. I: Scientific Reports. 2019 ; Bind 9, Nr. 1.
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title = "A method of predicting the in vitro fibril formation propensity of Abeta40 mutants based on their inclusion body levels in E. coli",
abstract = "Overexpression of recombinant proteins in bacteria may lead to their aggregation and deposition in inclusion bodies. Since the conformational properties of proteins in inclusion bodies exhibit many of the characteristics typical of amyloid fibrils. Based on these findings, we hypothesize that the rate at which proteins form amyloid fibrils may be predicted from their propensity to form inclusion bodies. To establish a method based on this concept, we first measured by SDS-PAGE and confocal microscopy the level of inclusion bodies in E. coli cells overexpressing the 40-residue amyloid-beta peptide, Aβ40, wild-type and 24 charge mutants. We then compared these results with a number of existing computational aggregation propensity predictors as well as the rates of aggregation measured in vitro for selected mutants. Our results show a strong correlation between the level of inclusion body formation and aggregation propensity, thus demonstrating the power of this approach and its value in identifying factors modulating aggregation kinetics.",
author = "Kalyani Sanagavarapu and Elisabeth N{\"u}ske and Irem Nasir and Georg Meisl and Immink, {Jasper N.} and Pietro Sormanni and Michele Vendruscolo and Knowles, {Tuomas P.J.} and Anders Malmendal and Celia Cabaleiro-Lago and Sara Linse",
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doi = "10.1038/s41598-019-39216-z",
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Sanagavarapu, K, Nüske, E, Nasir, I, Meisl, G, Immink, JN, Sormanni, P, Vendruscolo, M, Knowles, TPJ, Malmendal, A, Cabaleiro-Lago, C & Linse, S 2019, 'A method of predicting the in vitro fibril formation propensity of Abeta40 mutants based on their inclusion body levels in E. coli', Scientific Reports, bind 9, nr. 1, 3680. https://doi.org/10.1038/s41598-019-39216-z

A method of predicting the in vitro fibril formation propensity of Abeta40 mutants based on their inclusion body levels in E. coli. / Sanagavarapu, Kalyani; Nüske, Elisabeth; Nasir, Irem; Meisl, Georg; Immink, Jasper N.; Sormanni, Pietro; Vendruscolo, Michele; Knowles, Tuomas P.J.; Malmendal, Anders; Cabaleiro-Lago, Celia; Linse, Sara.

I: Scientific Reports, Bind 9, Nr. 1, 3680, 2019.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - A method of predicting the in vitro fibril formation propensity of Abeta40 mutants based on their inclusion body levels in E. coli

AU - Sanagavarapu, Kalyani

AU - Nüske, Elisabeth

AU - Nasir, Irem

AU - Meisl, Georg

AU - Immink, Jasper N.

AU - Sormanni, Pietro

AU - Vendruscolo, Michele

AU - Knowles, Tuomas P.J.

AU - Malmendal, Anders

AU - Cabaleiro-Lago, Celia

AU - Linse, Sara

PY - 2019

Y1 - 2019

N2 - Overexpression of recombinant proteins in bacteria may lead to their aggregation and deposition in inclusion bodies. Since the conformational properties of proteins in inclusion bodies exhibit many of the characteristics typical of amyloid fibrils. Based on these findings, we hypothesize that the rate at which proteins form amyloid fibrils may be predicted from their propensity to form inclusion bodies. To establish a method based on this concept, we first measured by SDS-PAGE and confocal microscopy the level of inclusion bodies in E. coli cells overexpressing the 40-residue amyloid-beta peptide, Aβ40, wild-type and 24 charge mutants. We then compared these results with a number of existing computational aggregation propensity predictors as well as the rates of aggregation measured in vitro for selected mutants. Our results show a strong correlation between the level of inclusion body formation and aggregation propensity, thus demonstrating the power of this approach and its value in identifying factors modulating aggregation kinetics.

AB - Overexpression of recombinant proteins in bacteria may lead to their aggregation and deposition in inclusion bodies. Since the conformational properties of proteins in inclusion bodies exhibit many of the characteristics typical of amyloid fibrils. Based on these findings, we hypothesize that the rate at which proteins form amyloid fibrils may be predicted from their propensity to form inclusion bodies. To establish a method based on this concept, we first measured by SDS-PAGE and confocal microscopy the level of inclusion bodies in E. coli cells overexpressing the 40-residue amyloid-beta peptide, Aβ40, wild-type and 24 charge mutants. We then compared these results with a number of existing computational aggregation propensity predictors as well as the rates of aggregation measured in vitro for selected mutants. Our results show a strong correlation between the level of inclusion body formation and aggregation propensity, thus demonstrating the power of this approach and its value in identifying factors modulating aggregation kinetics.

U2 - 10.1038/s41598-019-39216-z

DO - 10.1038/s41598-019-39216-z

M3 - Journal article

VL - 9

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

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ER -