title: Proteomic Identification and Analysis of Arginine-Methylated Proteins of Plasmodium falciparum at Asexual Blood Stages.
abstract: Plasmodium falciparum undergoes a tightly regulated developmental process in human erythrocytes, and recent studies suggest an important regulatory role of post-translational modifications (PTMs). As compared with Plasmodium phosphoproteome, little is known about other PTMs in the parasite. In the present study, we performed a global analysis of asexual blood stages of Plasmodium falciparum to identify arginine-methylated proteins. Using two different methyl arginine-specific antibodies, we immunoprecipitated the arginine-methylated proteins from the stage-specific parasite lysates and identified 843 putative arginine-methylated proteins by LC-MS/MS. Motif analysis of the protein sequences unveiled that the methylation sites are associated with the previously known methylation motifs such as GRx/RGx, RxG, GxxR, or WxxxR. We identified Plasmodium homologues of known arginine-methylated proteins in trypanosomes, yeast, and human. Hydrophilic interaction liquid chromatography (HILIC) was performed on the immunoprecipitates from the trophozoite stage to enrich arginine-methylated peptides. Mass spectrometry analysis of immunoprecipitated and HILIC fractions identified 55 arginine-methylated peptides having 62 methylated arginine sites. Functional classification revealed that the arginine-methylated proteins are involved in RNA metabolism, protein synthesis, intracellular protein trafficking, proteolysis, protein folding, chromatin organization, hemoglobin metabolic process, and several other functions. Summarily, the findings suggest that protein methylation of arginine residues is a widespread phenomenon in Plasmodium, and the PTM may play an important regulatory role in a diverse set of biological pathways, including host-pathogen interactions.
authors.affiliation: Malaria Biology Group, International Centre for Genetic Engineering and Biotechnology (ICGEB) , Aruna Asaf Ali Marg, New Delhi 110067, India.|Translational Bioinformatics Group, International Centre for Genetic Engineering and Biotechnology (ICGEB) , Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Zeeshan
authors.affiliation: Malaria Biology Group, International Centre for Genetic Engineering and Biotechnology (ICGEB) , Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Inderjeet
authors.initials: I
authors.lastname: Kaur
authors.affiliation: Translational Bioinformatics Group, International Centre for Genetic Engineering and Biotechnology (ICGEB) , Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Joseph
authors.initials: J
authors.lastname: Joy
authors.affiliation: Malaria Biology Group, International Centre for Genetic Engineering and Biotechnology (ICGEB) , Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Ekta
authors.initials: E
authors.lastname: Saini
authors.affiliation: Malaria Biology Group, International Centre for Genetic Engineering and Biotechnology (ICGEB) , Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Gourab
authors.initials: G
authors.lastname: Paul
authors.forename: Abhinav
authors.initials: A
authors.lastname: Kaushik
authors.affiliation: Malaria Biology Group, International Centre for Genetic Engineering and Biotechnology (ICGEB) , Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Surbhi
authors.initials: S
authors.lastname: Dabral
authors.affiliation: Parasite Cell Biology Group, International Centre for Genetic Engineering and Biotechnology (ICGEB) , Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Asif
authors.initials: A
authors.lastname: Mohmmed
authors.affiliation: Translational Bioinformatics Group, International Centre for Genetic Engineering and Biotechnology (ICGEB) , Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Dinesh
authors.initials: D
authors.lastname: Gupta
authors.affiliation: Malaria Biology Group, International Centre for Genetic Engineering and Biotechnology (ICGEB) , Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Pawan
authors.initials: P
authors.lastname: Malhotra
mesh_terms: D000595:Amino Acid Sequence; D001120:Arginine / Q000378:metabolism*; D002853:Chromatography, Liquid; D004912:Erythrocytes / Q000469:parasitology; D063990:Gene Ontology; D054884:Host-Pathogen Interactions; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D047468:Immunoprecipitation; D008018:Life Cycle Stages / Q000235:genetics; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D008745:Methylation; D058977:Molecular Sequence Annotation; D010963:Plasmodium falciparum / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D020543:Proteome; D015800:Protozoan Proteins; D001120:Arginine
keywords: LC−MS/MS; arginine methylation; malaria; parasite; plasmodium