Molecular cloning and demonstration of an aminopeptidase activity in a filarial nematode glycoprotein
ES-62 is an abundant phosphorylcholine-containing secreted glycoprotein of the filarial nematode Acanthocheilonema viteae. Using an antiserum directed against the parasite molecule, 3 cDNAs of size, ∼1.5–1.6 kbp were isolated from an A. viteae expression library. Sequence analysis in combination wit...
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Veröffentlicht in: | Molecular and biochemical parasitology 1999-10, Vol.104 (1), p.11-23 |
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Hauptverfasser: | , , , , , , , , , , , |
Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | ES-62 is an abundant phosphorylcholine-containing secreted glycoprotein of the filarial nematode
Acanthocheilonema viteae. Using an antiserum directed against the parasite molecule, 3 cDNAs of size, ∼1.5–1.6 kbp were isolated from an
A. viteae expression library. Sequence analysis in combination with N-terminal amino acid sequencing of purified ES-62 revealed that each clone contained a full-length cDNA for ES-62 corresponding to 474 amino acid residues but differed in their 5′ and 3′ untranslated regions. Characterisation of the 5′ end of ES-62 mRNA using 5′ rapid amplification of cDNA ends showed that it coded for a signal sequence. Several tryptic peptides were independently sequenced using quadruple-time-of-flight mass spectrometry and used to confirm the cDNA sequence. The mature protein was found to contain three potential N-linked glycosylation sites. Comparison of the derived amino acid sequence of ES-62 with the SwissProt database identified a sequence (between amino acid residues approximately 250 and 350 of mature ES-62) with significant similarity to several bacterial/fungal aminopeptidases. Incubation of ES-62 with leucine-7-amino-4-methylcoumarin as substrate confirmed that ES-62 possessed aminopeptidase activity. |
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ISSN: | 0166-6851 1872-9428 |
DOI: | 10.1016/S0166-6851(99)00113-9 |