This supplement requests a new Microwave Peptide Synthesizer for the parent project (R01- GM125853). The parent project proposed to study the molecular basis of membrane disruptions induced by the aggregation of -amyloid (A ) peptides. In order to achieve the research goals, a number of isotope- labeled A peptide sequences need to be synthesized to probe specific interactions between peptides and lipids, or to obtain molecular structural constraints to build structural models. This nature of the proposed research places peptide synthesis in a critical position. Currently, all peptide syntheses are completed manually in the PI?s laboratory. The manual synthesis is labor-intensive and time-consuming, and the crude products obtained usually contain relatively high abundances of impurities, which further cause lower yield after purification. The required instrument utilizes microwave, which significantly reduces the reaction time with more efficient and precise heating. By estimation, the synthesis time per peptide sequence can be reduced to ~ 20% and the byproducts in crude peptides can be largely eliminated. Therefore, the time allocation for peptide synthesis can be greatly shortened and the spectrometer time, such as the time spent on solid-state nuclear magnetic resonance (ssNMR) spectroscopy for full structural characterization, can be utilized more efficiently. For some aims, more quantitative methods will be applied, which requires high-purity peptides. The usage of microwave-based synthesis will help to achieve the reduction of side reactions. Overall, the requested instrument, if granted, will significantly benefit the proposed research in parent project. The synthesizer will be dedicated to the PI?s research project that is funded by NIGMS. A small portion of time, roughly 5%, will be used for other NIGMS-funded PIs who are either in the same department or collaborators to the PI. The equipment will be operated and maintained by the PI, who has ~ 20 years? experience in solid-phase peptide synthesis and the usage of peptide synthesizers. The Chemistry Department at Binghamton University also provides technical support in repairing scientific instrumentations with no charge for labors.
This project requests a microwave peptide synthesizer that will significantly benefit the proposed research activities in a NIGMS-funded project, which studies the molecular mechanisms of membrane disruptions induced by the aggregation of -amyloid (A ) peptides in Alzheimer?s disease (AD).
|Qiang, Wei; Doherty, Katelynne E (2018) Model Phospholipid Liposomes to Study the ?-Amyloid-Peptide-Induced Membrane Disruption. Methods Mol Biol 1777:355-367|
|Cheng, Qinghui; Hu, Zhi-Wen; Doherty, Katelynne E et al. (2018) The on-fibrillation-pathway membrane content leakage and off-fibrillation-pathway lipid mixing induced by 40-residue ?-amyloid peptides in biologically relevant model liposomes. Biochim Biophys Acta Biomembr :|