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Alzheimer's disease-associated Aβ42 peptide

May 19, 2017

Aggregation of the amyloid-beta peptide (Aβ) in the brain is strongly associated with Alzheimer´s disease (AD). This process is highly heterogeneous and transient in nature, thus hindering identification of the exact molecular form of Aβ responsible for the neurotoxicity observed in this disease. Therefore, characterizing Aβ aggregation is of utmost importance to make headway in the field of AD. Nuclear magnetic resonance spectroscopy (NMR), a technique used to obtain structural information, holds great potential to achieve this goal, as it could contribute to determining the structure of Aβ aggregate forms. However, it requires large amounts of peptide, as well as isotopic labels that are introduced through the Aβ peptide production process.

In this article, we report a new and straightforward production protocol to obtain the Aβ most strongly linked to AD, the so-called Aβ42 form, with the required labels for NMR experiments. Specifically, we describe an inexpensive strategy through which to obtain [U-15N]- and [U-2H,13C,15N]-labeled Aβ42. Notably, this approach does not require reversed phase high performance liquid chromatography (RP-HPLC), a costly and time-consuming purification technique widely used in previously reported Aβ production protocols. Instead, all the purification steps required in our production protocol can be performed with the fast protein liquid chromatography system (FPLC), which is widely available. The peptides that we obtained are of high purity and have the required isotope labeling to support NMR-based structural studies.Therefore, we conclude that this strategy offers a simpler and inexpensive approach to obtain isotopically labeled Aβ42 than previously described methods, thereby paving the way for NMR-based Aβ structural studies.
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For more information, please visit: http://www.eurekaselect.com/149791/article

Bentham Science Publishers

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