Identification of glycoproteins targeted by Trypanosoma cruzi trans-sialidase, a virulence factor that disturbs lymphocyte glycosylation.
Trypanosoma cruzi, the agent of the American trypanosomiasis or Chagas disease, bypasses its lack of de novo synthesis of sialic acids by expressing a surface-anchored trans-sialidase. This enzyme transfers sialic acid residues from the host's sialylglycoconjugates to the parasite's galactosylglycoconjugates. In addition to carrying out a pivotal role in parasite persistence/replication within the infected mammal, the trans-sialidase is shed into the bloodstream and induces alterations in the host immune system by modifying the sialylation of the immune cells. A major obstacle to understand these events is the difficulty to identify the transferred sialic acid among all those naturally occurring on the cell surface. Here, we report the use of azido-modified unnatural sialic acid to identify those molecules that act as cell surface acceptors of the sialyl residue in the trans-sialidase-catalyzed reaction, which might then be involved in the immune alterations induced. In living parasites, we readily observed the transfer of azido-sialic acid to surface mucins. When evaluating mouse thymocytes and splenocytes as acceptors of the azido-sugar, a complex pattern of efficiently tagged glycoproteins was revealed. In both leukocyte populations, the main proteins labeled were identified as different CD45 isoforms. Disruption of the cell architecture increased the number and the molecular weight distribution of azido-sialic acid tagged proteins. Nevertheless, CD45 remained to be the main acceptor. Mass spectrometry assays allowed us to identify other acceptors, mainly integrins. The findings reported here provide a molecular basis to understand the abnormalities induced in the immune system by the trans-sialidase during T. cruzi infection.
Muiá
Romina P
RP
Instituto de Investigaciones Biotecnológicas, Universidad Nacional de San Martín, B1650WGA San Martín, Argentina.
Yu
Hai
H
Prescher
Jennifer A
JA
Hellman
Ulf
U
Chen
Xi
X
Bertozzi
Carolyn R
CR
Campetella
Oscar
O
D000818:Animals; D006023:Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D006031:Glycosylation; D006790:Host-Parasite Interactions; D006801:Humans; D019169:Jurkat Cells; D008214:Lymphocytes / Q000378:metabolism*; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D009439:Neuraminidase / Q000737:chemistry* / Q000378:metabolism; D015800:Protozoan Proteins / Q000737:chemistry* / Q000378:metabolism; D014349:Trypanosoma cruzi / Q000201:enzymology* / Q000378:metabolism / Q000472:pathogenicity*; D037521:Virulence Factors / Q000737:chemistry* / Q000378:metabolism
D006023:Glycoproteins; D015800:Protozoan Proteins; D037521:Virulence Factors; C069715:trans-sialidase; D009439:Neuraminidase