Molecular Cloning, Primary Structure, and Properties of a New Glycoamidase from the Fungus Aspergillus tubigensis
| ISSN: | 0021-9258 |
|---|---|
| DOI: | 10.1074/jbc.272.36.22960 |
| Přístupová URL adresa: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::77a2a087fb2ab39ed2673aa9b1196cda https://doi.org/10.1074/jbc.272.36.22960 |
| Rights: | OPEN |
| Přírůstkové číslo: | edsair.doi.dedup.....77a2a087fb2ab39ed2673aa9b1196cda |
| Autor: | Nouzha Ftouhi-Paquin, Anthony L. Tarentino, Robert F. Stack, T.H. Plummer, Charles R. Hauer |
| Rok vydání: | 1997 |
| Předmět: |
Glycan
DNA Complementary Glycosylation Base Sequence Molecular Sequence Data Protein primary structure Cell Biology Biology Molecular cloning Peptide Mapping Biochemistry Fucose Amidase chemistry.chemical_compound Aspergillus chemistry Polysaccharides biology.protein Amino Acid Sequence Asparagine Cloning Molecular Molecular Biology Peptide sequence Chromatography High Pressure Liquid |
| Zdroj: | Journal of Biological Chemistry. 272:22960-22965 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.272.36.22960 |
| Popis: | A new glycoamidase, peptide-N4-(N-acetyl-beta-D-glucosaminyl)asparagine amidase (PNGase) At, was discovered in the eukaryote Aspergillus tubigensis. The enzyme was purified to homogeneity, and the DNA sequence was determined by cloning in Escherichia coli. Over 80% of the deduced amino acid sequence was verified independently by Edman analysis and/or electrospray ionization-mass spectrometry of protease fragments of native PNGase At. This glycoamidase contains 12 potential asparagine-linked glycosylation sites, of which at least 9 sites are occupied with typical high mannose oligosaccharides. PNGase At consists of two non-identical glycosylated subunits that are derived from a single polypeptide gene precursor. Evidence is presented suggesting that autocatalysis is involved in subunit formation. PNGase At is an important new tool for analysis of asparagine-linked glycans; it can hydrolyze a broad range of glycopeptides, including those with core-linked alpha1-->6 or alpha1-->3 fucose, under conditions not favorable with existing glycoamidases. |
| Databáze: | OpenAIRE |
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