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Phosphorylated Protein Levels in Animal-Sourced Food Muscles Based on Fe3+ and UV/Vis Spectrometry
journal contributionposted on 2022-02-18, 14:05 authored by Rong Cui, Mingke Shao, Hongyan Bi
Protein phosphorylation, a post-translational modification of proteins, is important in biological regulation. The quantity of phosphorylated proteins is a key requirement for the quality change of animal muscle foods. In the present study, a new approach to quantify phosphorylated proteins and/or peptides was developed based on ferric ions (Fe3+) and UV/vis spectrometry. This method is proved to be ultra-effective in discriminating phosphopeptides and non-phosphopeptides with the assistance of Fe3+. The protocol of extracting proteins with 0.1% trifluoroacetic acid (TFA) solution from animal muscle samples coupled with Fe3+ was verified by using an artificial mixture of peptides with different phosphorylation sites and was successfully used to characterize the phosphorylation quantity in the samples via UV/vis spectrometry. A peptide with one phosphorylated site was taken as a reference standard and successfully utilized for the absolute quantification of phosphorylated proteins in caprine muscles during frozen storage and in fish muscle food samples. This present study paves a new way for the evaluation of phosphorylated protein quantitative levels in bio-samples.
phosphorylated protein levelsone phosphorylated sitedifferent phosphorylation sitespresent study pavesanimal muscle foods3 + supquantify phosphorylated proteinssamples via uvpresent studyphosphorylated proteinsvis spectrometrytrifluoroacetic acidtranslational modificationsuccessfully utilizedsuccessfully usedreference standardquality changephosphorylation quantitynew waynew approachkey requirementfrozen storageferric ionsextracting proteinsdeveloped basedcaprine musclesbiological regulationartificial mixtureabsolute quantification