Schistosoma mansoni: Sm23 is a transmembrane protein that also contains a glycosylphosphatidylinositol anchor.
Sm23, a surface protein of the human parasite Schistosoma mansoni, belongs to the family of "cysteine-rich, hydrophobic proteins," which are expressed on mammalian hematopoietic cells or tumor cells. Sm23 shares the highly conserved hydrophobicity profile of these proteins, which predicts four transmembrane segments, but is in addition linked to the membrane by a glycosylphosphatidylinositol (GPI) anchor. Our results suggest that Sm23 uses both the potential transmembrane domains and the GPI anchor for membrane insertion: (a) Sm23 was not released from the surface after cleavage with phosphatidylinositol-specific phospholipase C (PIPLC). (b) In a Triton X-114 phase-separation system, native [3H]ethanolamine- or [35S]methionine-labeled Sm23 partitioned into the detergent phase. Upon removal of the GPI anchor by PIPLC, the majority of the molecules stayed in the detergent-phase as expected of a transmembrane protein. (c) When full-length recombinant Sm23 was transcribed and translated in vitro, the polypeptide chain was inserted into microsomal membranes: Sm23 stayed associated with the membranes when they were incubated with carbonate buffer at pH 11.5, and membrane bound Sm23 was protected from digestion with proteinase K. (d) Recombinant Sm23, when expressed in the baculovirus expression system, was transported to the surface of infected insect cells, and similarly to the native protein it was not released from these cells after cleavage with PIPLC.
Köster
B
B
Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Strand
M
M
D000818:Animals; D000947:Antigens, Helminth / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016367:Baculoviridae / Q000235:genetics; D001692:Biological Transport; D002474:Cell-Free System; D017261:Glycosylphosphatidylinositols / Q000032:analysis*; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D009036:Moths / Q000166:cytology / Q000378:metabolism; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D054801:Phosphoinositide Phospholipase C; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000378:metabolism; D012156:Reticulocytes; D012550:Schistosoma mansoni / Q000737:chemistry*
D000947:Antigens, Helminth; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; D011994:Recombinant Proteins; C067614:antigen Sm23, Schistosoma; D010727:Phosphoric Diester Hydrolases; D054801:Phosphoinositide Phospholipase C; D043265:Phosphatidylinositol Diacylglycerol-Lyase