GPIomics: global analysis of glycosylphosphatidylinositol-anchored molecules of Trypanosoma cruzi.
Glycosylphosphatidylinositol (GPI) anchoring is a common, relevant posttranslational modification of eukaryotic surface proteins. Here, we developed a fast, simple, and highly sensitive (high attomole-low femtomole range) method that uses liquid chromatography-tandem mass spectrometry (LC-MS(n)) for the first large-scale analysis of GPI-anchored molecules (i.e., the GPIome) of a eukaryote, Trypanosoma cruzi, the etiologic agent of Chagas disease. Our genome-wise prediction analysis revealed that approximately 12% of T. cruzi genes possibly encode GPI-anchored proteins. By analyzing the GPIome of T. cruzi insect-dwelling epimastigote stage using LC-MS(n), we identified 90 GPI species, of which 79 were novel. Moreover, we determined that mucins coded by the T. cruzi small mucin-like gene (TcSMUG S) family are the major GPI-anchored proteins expressed on the epimastigote cell surface. TcSMUG S mucin mature sequences are short (56-85 amino acids) and highly O-glycosylated, and contain few proteolytic sites, therefore, less likely susceptible to proteases of the midgut of the insect vector. We propose that our approach could be used for the high throughput GPIomic analysis of other lower and higher eukaryotes.
Department of Biological Sciences, The Border Biomedical Research Center, University of Texas at El Paso, El Paso, TX 79968, USA.
Ernesto S
ES
Nakayasu
Dmitry V
DV
Yashunsky
Lilian L
LL
Nohara
Ana Claudia T
AC
Torrecilhas
Andrei V
AV
Nikolaev
Igor C
IC
Almeida
D000595:Amino Acid Sequence; D000818:Animals; D002853:Chromatography, Liquid; D019286:Endopeptidase K / Q000378:metabolism; D018503:Genome, Protozoan; D054794:Glycomics; D017261:Glycosylphosphatidylinositols / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D008055:Lipids / Q000032:analysis; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D011134:Polysaccharides / Q000032:analysis; D011137:Polystyrenes; D015800:Protozoan Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013997:Time Factors; D014349:Trypanosoma cruzi / Q000235:genetics / Q000378:metabolism*; D014357:Trypsin / Q000378:metabolism
D017261:Glycosylphosphatidylinositols; D008055:Lipids; D010455:Peptides; D011134:Polysaccharides; D011137:Polystyrenes; D015800:Protozoan Proteins; D014357:Trypsin; D019286:Endopeptidase K