Department of Chemistry and Biochemistry, Kettering University, Flint, Michigan, United States of America.
PLoS One. 2021 Mar 22;16(3):e0248717. doi: 10.1371/journal.pone.0248717. eCollection 2021.
Wheat germ acid phosphatase (WGAP) is a commercial preparation of partially purified protein commonly used in laboratory settings for non-specific enzymatic dephosphorylation. It is known that these preparations contain multiple phosphatase isozymes and are still relatively crude. This study therefore aimed to identify the protein components of a commercial preparation of wheat germ acid phosphatase using mass spectroscopy and comparative genomics. After one post-purchase purification step, the most prevalent fifteen proteins in the mixture included heat shock proteins, beta-amylases, glucoseribitol dehydrogenases, enolases, and an aminopeptidase. While not among the most abundant components, eight unique dephosphorylation enzymes were also present including three purple acid phosphatases. Furthermore, it is shown that some of these correspond to previously isolated isozymes; one of which has been also previously shown by transcriptome data to be overexpressed in wheat seeds. In summary, this study identified the major components of WGAP including phosphatases and hypothesizes the most active components towards a better understanding of this commonly used laboratory tool.
小麦胚芽酸性磷酸酶(WGAP)是一种经过部分纯化的蛋白质商业制剂,常用于实验室中非特异性酶去磷酸化。已知这些制剂含有多种磷酸酶同工酶,仍然相对粗糙。因此,本研究旨在使用质谱和比较基因组学鉴定小麦胚芽酸性磷酸酶商业制剂的蛋白质成分。在购买后进行一次纯化步骤后,混合物中最常见的十五种蛋白质包括热休克蛋白、β-淀粉酶、葡萄糖山梨醇脱氢酶、烯醇酶和氨肽酶。虽然不是最丰富的成分之一,但也存在八种独特的去磷酸化酶,包括三种紫色酸性磷酸酶。此外,研究表明其中一些与先前分离的同工酶相对应;其中一种同工酶之前也通过转录组数据表明在小麦种子中过表达。总之,本研究鉴定了 WGAP 的主要成分,包括磷酸酶,并假设最活跃的成分有助于更好地理解这种常用的实验室工具。