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Amyloid precursor protein mutation E682K at the alternative β-secretase cleavage β'-site increases Aβ generation.

Zhou L, Brouwers N, Benilova I, Vandersteen A, Mercken M, Van Laere K, Van Damme P, Demedts D, Van Leuven F, Sleegers K, Broersen K, Van Broeckhoven C, Vandenberghe R, De Strooper B - EMBO Mol Med (2011)

Bottom Line: Increasing exon- and exome-based sequencing efforts will identify many more putative pathogenic mutations without conclusive segregation-based evidence in a single family.Our study shows how functional analysis of such mutations allows to determine the potential pathogenic nature of these mutations.We propose to classify the E682K mutation as probable pathogenic awaiting further independent confirmation of its association with AD in other patients.

View Article: PubMed Central - PubMed

Affiliation: Department for Developmental and Molecular Genetics, VIB, Leuven, Belgium.

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Related in: MedlinePlus

Biophysical determination of the aggregation kinetics of WT Aβ 42 and mutant E11K Aβ42ThT fluorescence of aggregating WT Aβ42 (black) and mutant Aβ42 E11K (red). Indicated are the error bars for each time point based on three measurements. The aggregation kinetics is not dramatically affected by the E11K mutation, apart from a small increase in the initial rate during the first 3 h of aggregation process.TEM images further underline that the aggregation process of Aβ42 upon introduction of the E11K mutation is not significantly modified. After 1.5 h aggregates are observed which become larger upon further incubation.
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fig05: Biophysical determination of the aggregation kinetics of WT Aβ 42 and mutant E11K Aβ42ThT fluorescence of aggregating WT Aβ42 (black) and mutant Aβ42 E11K (red). Indicated are the error bars for each time point based on three measurements. The aggregation kinetics is not dramatically affected by the E11K mutation, apart from a small increase in the initial rate during the first 3 h of aggregation process.TEM images further underline that the aggregation process of Aβ42 upon introduction of the E11K mutation is not significantly modified. After 1.5 h aggregates are observed which become larger upon further incubation.

Mentions: We studied the in vitro aggregation properties of synthetic Aβ peptide carrying the E682K mutation (E11K mutant Aβ). The aggregation kinetics of WT and mutant Aβ42 peptide were monitored by a Thioflavin T (ThT) fluorescence assay. Compared with WT Aβ42 peptide, E11K mutant Aβ42 showed a slight increase in the initial aggregation rate during the first 3 h of the aggregation process (Fig 5A); however, the overall change in aggregation kinetics was small. Transmission electron microscopy (TEM) images further underlined that the aggregation process of E11K mutant Aβ42 was not significantly different from that of the WT Aβ42 peptide (Fig 5B). In parallel, we also analysed the cytotoxicity of E11K mutant and WT Aβ42, no significant difference was detected (Fig S2 of Supporting Information).


Amyloid precursor protein mutation E682K at the alternative β-secretase cleavage β'-site increases Aβ generation.

Zhou L, Brouwers N, Benilova I, Vandersteen A, Mercken M, Van Laere K, Van Damme P, Demedts D, Van Leuven F, Sleegers K, Broersen K, Van Broeckhoven C, Vandenberghe R, De Strooper B - EMBO Mol Med (2011)

Biophysical determination of the aggregation kinetics of WT Aβ 42 and mutant E11K Aβ42ThT fluorescence of aggregating WT Aβ42 (black) and mutant Aβ42 E11K (red). Indicated are the error bars for each time point based on three measurements. The aggregation kinetics is not dramatically affected by the E11K mutation, apart from a small increase in the initial rate during the first 3 h of aggregation process.TEM images further underline that the aggregation process of Aβ42 upon introduction of the E11K mutation is not significantly modified. After 1.5 h aggregates are observed which become larger upon further incubation.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3377078&req=5

fig05: Biophysical determination of the aggregation kinetics of WT Aβ 42 and mutant E11K Aβ42ThT fluorescence of aggregating WT Aβ42 (black) and mutant Aβ42 E11K (red). Indicated are the error bars for each time point based on three measurements. The aggregation kinetics is not dramatically affected by the E11K mutation, apart from a small increase in the initial rate during the first 3 h of aggregation process.TEM images further underline that the aggregation process of Aβ42 upon introduction of the E11K mutation is not significantly modified. After 1.5 h aggregates are observed which become larger upon further incubation.
Mentions: We studied the in vitro aggregation properties of synthetic Aβ peptide carrying the E682K mutation (E11K mutant Aβ). The aggregation kinetics of WT and mutant Aβ42 peptide were monitored by a Thioflavin T (ThT) fluorescence assay. Compared with WT Aβ42 peptide, E11K mutant Aβ42 showed a slight increase in the initial aggregation rate during the first 3 h of the aggregation process (Fig 5A); however, the overall change in aggregation kinetics was small. Transmission electron microscopy (TEM) images further underlined that the aggregation process of E11K mutant Aβ42 was not significantly different from that of the WT Aβ42 peptide (Fig 5B). In parallel, we also analysed the cytotoxicity of E11K mutant and WT Aβ42, no significant difference was detected (Fig S2 of Supporting Information).

Bottom Line: Increasing exon- and exome-based sequencing efforts will identify many more putative pathogenic mutations without conclusive segregation-based evidence in a single family.Our study shows how functional analysis of such mutations allows to determine the potential pathogenic nature of these mutations.We propose to classify the E682K mutation as probable pathogenic awaiting further independent confirmation of its association with AD in other patients.

View Article: PubMed Central - PubMed

Affiliation: Department for Developmental and Molecular Genetics, VIB, Leuven, Belgium.

Show MeSH
Related in: MedlinePlus