一种表达和纯化Aβ（M1-42）的高效方法。
An Efficient Method for the Expression and Purification of Aβ(M1-42).
作者信息
Yoo Stan, Zhang Sheng, Kreutzer Adam G, Nowick James S
机构信息
Department of Chemistry , University of California, Irvine , Irvine , California 92697-2025 , United States.
出版信息
Biochemistry. 2018 Jul 3;57(26):3861-3866. doi: 10.1021/acs.biochem.8b00393. Epub 2018 May 24.
Advances in amyloid research rely on improved access to the β-amyloid peptide, Aβ. N-Terminal methionine-extended Aβ, Aβ(M1-42), is a readily expressed and widely used form of Aβ with properties comparable to those of the natural Aβ(1-42) peptide. Expression of Aβ(M1-42) is simple to execute and avoids an expensive and often difficult enzymatic cleavage step associated with expression and isolation of Aβ(1-42). This paper reports an efficient method for the expression and purification of Aβ(M1-42) and N-labeled Aβ(M1-42). This method affords the pure peptide at ∼19 mg/L of bacterial culture through simple and inexpensive steps in 3 days. This paper also reports a simple method for the construction of recombinant plasmids and the expression and purification of Aβ(M1-42) peptides containing familial mutations. We anticipate that these methods will enable experiments that would otherwise be hindered by insufficient access to Aβ.
淀粉样蛋白研究的进展依赖于对β-淀粉样肽（Aβ）更便捷的获取。N端甲硫氨酸延伸的Aβ，即Aβ（M1-42），是一种易于表达且广泛使用的Aβ形式，其性质与天然Aβ（1-42）肽相当。Aβ（M1-42）的表达操作简单，避免了与Aβ（1-42）表达和分离相关的昂贵且通常困难的酶切步骤。本文报道了一种高效表达和纯化Aβ（M1-42）以及N标记的Aβ（M1-42）的方法。该方法通过简单且廉价的步骤，在3天内从每升细菌培养物中获得约19毫克的纯肽。本文还报道了一种构建重组质粒以及表达和纯化含家族性突变的Aβ（M1-42）肽的简单方法。我们预计这些方法将使那些因无法充分获取Aβ而受阻的实验得以开展。
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