Abstract
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.
Highlights
Phosphatases catalyze the hydrolysis of phosphoric-monoester compounds to produce a free inorganic phosphate and an alcohol
While the name wheat germ acid phosphatase implies a singular protein, it typically refers to a commercial preparation of partially purified protein which is known to contain multiple proteins [13,14,15,16,17,18]
The annotated genomes from four wheat species (Triticum aestivum, Triticum dicoccoides, Triticum urartu, and Aegilops tauschii) were investigated in the present study
Summary
Phosphatases catalyze the hydrolysis of phosphoric-monoester compounds to produce a free inorganic phosphate and an alcohol. Purple acid phosphatases (PAPs), for example, are metalloenzymes that appear purple or violet in their oxidized form. Commercial wheat germ acid phosphatase (WGAP) is extracted from the germ/embryo of wheat plants and is commonly used in vitro in many biochemistry laboratories. Its availability and inexpensive cost have historically made it a common choice for many projects which require a broad specificity phosphatase, including those in education and research (see, for example, references [2,3,4,5,6,7,8,9,10,11,12]). While the name wheat germ acid phosphatase implies a singular protein, it typically refers to a commercial preparation of partially purified protein which is known to contain multiple proteins [13,14,15,16,17,18].
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