Characterization of a novel GDP-mannose : Serine-protein mannose-l-phosphotransferase from Leishmania mexicana
- Author(s)
- Moss, JM; Reid, GE; Mullin, KA; Zawadzki, JL; Simpson, RJ; McConville, MJ;
- Details
- Publication Year 1999-03-05,Volume 274,Issue #10,Page 6678-6688
- Journal Title
- JOURNAL OF BIOLOGICAL CHEMISTRY
- Publication Type
- Journal Article
- Abstract
- Protozoan parasites of the genus Leishmania secrete a number of glycoproteins and mucin-like proteoglycans that appear to be important parasite virulence factors. We have previously proposed that the polypeptide backbones of these molecules are extensively modified with a complex array of phosphoglycan chains that are linked to Ser/Thr-rich domains via a common Man alpha 1-PO4-Ser Linkage (Ilg, T., Overath, P., Ferguson, M. A J., Rutherford, T,, Campbell, D, G,, and McConville, M, J, (1994) J, Biol. Chem, 269, 24073-24081), In this study, we show that Leishmania mexicana promastigotes contain a peptide-specific mannose-l-phosphotransferase (pep-MPT) activity that adds Man alpha 1-P to serine residues in a range of defined peptides, The presence and location of the Man alpha 1-PO4-Ser linkage in these peptides were determined by electrospray ionization mass spectrometry and chemical and enzymatic treatments. The pep-MPT activity was solubilized in non-ionic detergents, was dependent on Mn2+, utilized GDP-Man as the mannose donor, and was expressed in all developmental stages of the parasite. The pep-MPT activity was maximal against peptides containing Ser/Thr-rich domains of the endogenous accepters and, based on competition assays with oligosaccharide accepters, was distinct from other leishmanial MPTs involved in the initiation and elongation of lipid-linked phosphoglycan chains. In subcellular fractionation experiments, pep-MPT was resolved from the endoplasmic reticulum marker BiP, but had an overlapping distribution with the cis-Golgi marker Rab1, Although Man-PO4 residues in the mature secreted glycoproteins are extensively modified with mannose oligosaccharides and phosphoglycan chains, similar modifications were not added to peptide-linked Man-PO4 residues in the in vitro assays, Similarly, Man-PO4 residues on endogenous polypeptide accepters were also poorly extended, although the elongating enzymes were still active, suggesting that the pep-MPT activity and elongating enzymes may be present in separate subcellular compartments.
- Publisher
- AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
- Keywords
- SECRETED ACID-PHOSPHATASE; LIPOPHOSPHOGLYCAN; BIOSYNTHESIS; PROTEOPHOSPHOGLYCAN; AMASTIGOTES; PARASITES; PROMASTIGOTES; PURIFICATION; DONOVANI; EPITOPES
- Publisher's Version
- https://doi.org/10.1074/jbc.274.10.6678
- Terms of Use/Rights Notice
- Refer to copyright notice on published article.
Creation Date: 1999-03-05 12:00:00