Trypanosoma congolense procyclins: unmasking cryptic major surface glycoproteins in procyclic forms.
In the tsetse fly, the protozoan parasite Trypanosoma congolense is covered by a dense layer of glycosylphosphatidylinositol (GPI)-anchored molecules. These include a protease-resistant surface molecule (PRS), which is expressed by procyclic forms early in infection, and a glutamic acid- and alanine-rich protein (GARP), which appears at later stages. Since neither of these surface antigens is expressed at intermediate stages, we investigated whether a GPI-anchored protein of 50 to 58 kDa, previously detected in procyclic culture forms, might constitute the coat of these parasites. We therefore partially purified the protein from T. congolense Kilifi procyclic forms, obtained an N-terminal amino acid sequence, and identified its gene. Detailed analyses showed that the mature protein consists almost exclusively of 13 heptapeptide repeats (EPGENGT). The protein is densely N glycosylated, with up to 13 high-mannose oligosaccharides ranging from Man(5)GlcNAc(2) to Man(9)GlcNAc(2) linked to the peptide repeats. The lipid moiety of the glycosylphosphatidylinositol is composed of sn-1-stearoyl-2-lyso-glycerol-3-HPO(4)-1-(2-O-acyl)-d-myo-inositol. Heavily glycosylated proteins with similar repeats were subsequently identified in T. congolense Savannah procyclic forms. Collectively, this group of proteins was named T. congolense procyclins to reflect their relationship to the EP and GPEET procyclins of T. brucei. Using an antiserum raised against the EPGENGT repeat, we show that T. congolense procyclins are expressed continuously in the fly midgut and thus form the surface coat of cells that are negative for both PRS and GARP.
Utz
Silvia
S
Institute of Biochemistry & Molecular Medicine, University of Bern, Bern, Switzerland.
Roditi
Isabel
I
Kunz Renggli
Christina
C
Almeida
Igor C
IC
Acosta-Serrano
Alvaro
A
Bütikofer
Peter
P
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D018076:DNA, Complementary; D008055:Lipids / Q000737:chemistry; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D019151:Peptide Library; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020449:Repetitive Sequences, Amino Acid; D016415:Sequence Alignment; D021241:Spectrometry, Mass, Electrospray Ionization; D014348:Trypanosoma congolense / Q000737:chemistry* / Q000378:metabolism; D014370:Tsetse Flies / Q000469:parasitology
D018076:DNA, Complementary; D008055:Lipids; D008562:Membrane Glycoproteins; D019151:Peptide Library; D015800:Protozoan Proteins; C053277:procyclic acidic repetitive protein, Trypanosoma