Effect of the Histone Lysine Acetyltransferase MYST-A on the Protein Post-translational Modifications of Toxoplasma gondii
The aim of this study was to investigate the effect of the histone lysine acetyltransferase MYST-A on the protein post-translational modifications of Toxoplasma gondii.The full-length gene sequence was obtained from ToxoDB database of Toxoplasma gondii,the expression vector was constructed,the recombinant protein of His tag was purified in large quantities,and the experimental animals were immunized,polyclonal antibodies were prepared and purified.The subcellular localization of Toxoplasma gondii was analyzed by indirect immunofluorescence assay,and different concentrations of antibodies were used to act on Toxoplasma gondii and the whole parasite protein was extracted for Western-blot verification to analyze whether it had an effect on the protein post-translational modifications of Toxoplasma gondii.ALP2a protein is an important component of histone acetyltransferase complex.MYST-A and ALP2a may form a histone acetyltransferase complex to participate in post-translational modifications of histones.This study intends to verify whether they interact with each other through molecular interaction experiments.The results showed that the recombinant expression vector was successfully constructed,and the recombinant protein His-MYST-A were purified.His-MYST-A immunized mice and rabbits and produced specific polyclonal antibodies.The results of IFA analysis showed that MYST-A was widely distributed in the cytoplasm of Toxoplasma gondii both in the vacuoles and in the tachyzoites.The effect of MYST-A on the protein post-translational modifications of Toxoplasma gondii showed that with the increase of antibody concentration,the levels of ubiquitination,monomethyl/dimethylation,malonylation and succinylation increase(especially at 55 kDa),indicating that the protein played a negative regulatory role in them.The interaction between MYST-A and ALP2a was verified by molecular interaction experiment.This study further revealed that the histone lysine acetyltransferase MYST-A of Toxoplasma gondii plays an important role in the regulation of protein post-translational modifications,which lays a foundation for further study of its biological functions.The study of protein post-translational modifications is great significance to understand the pathogenic mechanism of Toxoplasma gondii and its interaction with the host.It also provides some reference value and theoretical basis for the development of novel Toxoplasma gondii vaccines and the screening of drug targets.
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