Detection of beta-amyloid peptide aggregation using DNA electrophoresis.
DNA could readily associate with the aggregated forms of the beta-amyloid peptides beta(1-40) and beta(25-35), giving rise to a shift in the electrophoretic mobility of DNA. As a result, DNA was retained at the top of a 1% agarose gel. In contrast, the electrophoretic mobility of DNA was little influenced by the monomeric forms of beta(1-40) and beta(25-30). DNA from different sources such as lambda phage, Escherichia coli plasmid, and human gene showed similar results. However, the electrophoretic mobility of RNA was shifted by the monomeric beta(1-40) and beta(25-35) as well as by the aggregated beta(1-40) and beta(25-35). The association of DNA with the aggregated beta-amyloid peptides could occur at pH 4-9. The inhibitory action of hemin on beta-amyloid aggregation could be confirmed using the DNA mobility shift assay. These results indicate that the DNA mobility shift assay is useful for kinetic study of beta-amyloid aggregation as well as for testing of agents that might modulate beta-amyloid aggregation.
Ahn
B W
BW
Chonnam National University Research Institute of Medical Sciences, Hakdong 5, Donggu, Kwangju, 501-190, Korea. bwahn@chonnam.ac.kr
Song
D U
DU
Jung
Y D
YD
Chay
K O
KO
Chung
M A
MA
Yang
S Y
SY
Shin
B A
BA
D016229:Amyloid beta-Peptides / Q000378:metabolism*; D015342:DNA Probes; D004121:Dimethyl Sulfoxide; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D006801:Humans; D010446:Peptide Fragments / Q000378:metabolism*; D011485:Protein Binding
D016229:Amyloid beta-Peptides; D015342:DNA Probes; D010446:Peptide Fragments; C078592:amyloid beta-protein (1-40); C074636:amyloid beta-protein (25-35); D004121:Dimethyl Sulfoxide