Structural analysis and antibody response to the extracellular glutathione S-transferases from Onchocerca volvulus.
Onchocerca volvulus is a human pathogenic filarial parasite which, like other parasitic nematodes, is capable of surviving in an immunologically competent host by employing a variety of immune evasion strategies and defense mechanisms including the detoxification and repair mechanisms of the glutathione S-transferases (GSTs). In this study we analyzed the glycosylation pattern and the immunological properties of extracellular O. volvulus GST1a and -1b (OvGST1a and -1b). The enzymes differ in only 10 amino acids, and both are glycoproteins that have cleavable signal peptides and unusual N-terminal extensions. These characteristics have not been described for other GSTs so far. Mass spectrometry analyses indicate that both enzymes carry high-mannose type oligosaccharides on at least four glycosylation sites. Glycosylation sites 1 to 3 of OvGST1a (OvGST1b sites 2 to 4) are occupied by truncated N-glycans (Man(2)GlcNAc2 to Man(5)GlcNAc(2)), and N glycosylation site 4 of OvGST1a (OvGST1b site 5) carries Man(5)GlcNAc2 to Man(9)GlcNAc(2). To analyze the capacity of these secretory GSTs to stimulate host immune responses, we studied the antibody responses of onchocerciasis patients against the native affinity-purified OvGST1a and -1b. By enzyme-linked immunosorbent assay we showed that OvGST1a and -1b are immunodominant antigens, with less than 7% nonresponder patients. A direct comparison of the antibody responses to the glycosylated and deglycosylated forms demonstrates the high immunogenicity of the N-glycans. Analyses of the antibody responses to the unusual N-terminal extension show an enhanced recognition of this portion by patients as opposed to recognition of the recombinant protein without extension.
Sommer
A
A
Bernhard Nocht Institute for Tropical Medicine, 20359 Hamburg, Germany.
Nimtz
M
M
Conradt
H S
HS
Brattig
N
N
Boettcher
K
K
Fischer
P
P
Walter
R D
RD
Liebau
E
E
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood*; D000947:Antigens, Helminth / Q000737:chemistry / Q000276:immunology*; D002240:Carbohydrate Sequence; D005260:Female; D005982:Glutathione Transferase / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000302:isolation & purification; D017181:Onchocerca volvulus / Q000201:enzymology / Q000276:immunology*; D009855:Onchocerciasis / Q000276:immunology* / Q000469:parasitology; D011498:Protein Precursors / Q000276:immunology; D021382:Protein Sorting Signals / Q000502:physiology
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D006023:Glycoproteins; D007527:Isoenzymes; D009844:Oligosaccharides; D011498:Protein Precursors; D021382:Protein Sorting Signals; D005982:Glutathione Transferase