Molecular cloning and characterization of a novel repeat-containing Leishmania major gene, ppg1, that encodes a membrane-associated form of proteophosphoglycan with a putative glycosylphosphatidylinositol anchor.
Leishmania parasites secrete a variety of proteins that are modified by phosphoglycan chains structurally similar to those of the cell surface glycolipid lipophosphoglycan. These proteins are collectively called proteophosphoglycans. We report here the cloning and sequencing of a novel Leishmania major proteophosphoglycan gene, ppg1. It encodes a large polypeptide of approximately 2300 amino acids. The N-terminal domain of approximately 70 kDa exhibits 11 imperfect amino acid repeats that show some homology to promastigote surface glycoproteins of the psa2/gp46 complex. The large central domain apparently consists exclusively of approximately 100 repetitive peptides of the sequence APSASSSSA(P/S)SSSSS(+/-S). Gene fusion experiments demonstrate that these peptide repeats are the targets of phosphoglycosylation in Leishmania and that they form extended filamentous structures reminiscent of mammalian mucins. The C-terminal domain contains a functional glycosylphosphatidylinositol anchor addition signal sequence, which confers cell surface localization to a normally secreted Leishmania acid phosphatase, when fused to its C terminus. Antibody binding studies show that the ppg1 gene product is phosphoglycosylated by phosphoglycan repeats and cap oligosaccharides. In contrast to previously characterized proteophosphoglycans, the ppg1 gene product is predominantly membrane-associated and it is expressed on the promastigote cell surface. Therefore this membrane-bound proteophosphoglycan may be important for direct host-parasite interactions.
Ilg
T
T
Max-Planck-Institut für Biologie, Abteilung Membranbiochemie, Corrensstrasse 38, D-72076 Tübingen, Germany. thomas.ilg@tuebingen.mpg.de
Montgomery
J
J
Stierhof
Y D
YD
Handman
E
E
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017125:Genes, Protozoan; D015698:Genomic Library; D006028:Glycosphingolipids / Q000235:genetics; D006031:Glycosylation; D017261:Glycosylphosphatidylinositols; D006790:Host-Parasite Interactions; D018320:Leishmania major / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D021382:Protein Sorting Signals; D011509:Proteoglycans / Q000235:genetics* / Q000648:ultrastructure; D015800:Protozoan Proteins; D020449:Repetitive Sequences, Amino Acid; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid
D006028:Glycosphingolipids; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; C400527:Ppg1 protein, Leishmania; D021382:Protein Sorting Signals; D011509:Proteoglycans; D015800:Protozoan Proteins; C008290:lipophosphonoglycan