Synthesis of Abeta[1-42] and its derivatives with improved efficiency.
It has been proved that the principal component of senile plaques is aggregates of beta-amyloid peptide (Abeta) in cases of one of the most common forms of age-related neurodegenerative disorders, Alzheimer's disease (AD). Although the synthetic methods for the synthesis of Abeta peptides have been developed since their first syntheses, Abeta[1-42] is still problematic to prepare. The highly hydrophobic composition of Abeta[1-42] results in aggregation between resin-bound peptide chains or intrachain aggregation which leads to a decrease in the rates of deprotection and repetitive incomplete coupling reactions during 9-flurenylmethoxycarbonyl (Fmoc) synthesis. In order to avoid aggregation and/or disrupt internal aggregation during stepwise Fmoc solid phase synthesis and to improve the quality of crude products, several attempts have been made. Since highly pure Abeta peptides in large quantities are used in biological experiments, we wanted to develop a method for a rational synthesis of human Abeta[1-42] with high purity and adequate yield. This paper reports a convenient methodology with a novel solvent system for the synthesis of Abeta[1-42], its N-terminally truncated derivatives Abeta[4-42] and Abeta[5-42], and Abeta[1-42] labeled with 7-amino-4-methyl-3-coumarinylacetic acid (AMCA) at the N-terminus using Fmoc strategy. The use of 10% anisole in Dimethylformamide/Dichloromethane (DMF/DCM) can substantially improve the purity and yield of crude Abeta[1-42] and has been shown to be an optimal coupling condition for the synthesis of Abeta[1-42]. Anisole is a cheap and simple aid in the synthesis of 'difficult sequences' where other solvents are less successful in the prevention of aggregation during the synthesis.
Zarándi
Márta
M
Department of Medical Chemistry, University of Szeged, H-6720 Szeged, Dóm tér 8, Hungary. zarandimarta@yahoo.com
Soós
Katalin
K
Fülöp
Lívia
L
Bozsó
Zsolt
Z
Datki
Zsolt
Z
Tóth
Gábor K
GK
Penke
Botond
B
D000595:Amino Acid Sequence; D016229:Amyloid beta-Peptides / Q000138:chemical synthesis* / Q000302:isolation & purification / Q000633:toxicity; D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes; D008969:Molecular Sequence Data; D009498:Neurotoxins; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000302:isolation & purification / Q000633:toxicity
D016229:Amyloid beta-Peptides; D005456:Fluorescent Dyes; D009498:Neurotoxins; D010446:Peptide Fragments; C075222:amyloid beta-protein (1-42)