Open AccessThe Glycosylation of the Aspartic Proteinases from Barley ( Hordeum Vulgare L.) and Cardoon ( Cynara Cardunculus L.)
Author(s) -
Costa Júlia,
Ashford David A.,
Nimtz Manfred,
Bento Isabel,
FrazÃo Carlos,
Esteves Cristina L.,
Faro Carlos J.,
Kervinen Jukka,
Pires Euclides,
VerÍssimo Paula,
Wlodawer Alexander,
Carrondo Maria Arménia
Publication year - 1997
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1997.t01-1-00695.x
Subject(s) - cynara , exoglycosidase , glycosylation , hordeum vulgare , glycan , biochemistry , chemistry , n linked glycosylation , proteinase 3 , biology , botany , glycoprotein , poaceae , inflammation , myeloperoxidase , immunology
Plant aspartic proteinases characterised at the molecular level contain one or more consensus N‐glycosylation sites [Runeberg‐Roos, P., Törmäkangas, K. & Östman, A. (1991) Eur. J. Biochem. 202 , 1021–1027; Asakura, T., Watanabe, H., Abe, K. & Arai, S. (1995) Eur. J. Biochem. 232 , 77–83; Veríssimo, P., Faro, C., Moir, A. J. G., Lin, Y., Tang, J. & Pires, E. (1996) Eur. J. Biochem. 235 , 762–7681. We found that the glycosylation sites are occupied for the barley ( Hordeum vulgare L.) aspartic proteinase (Asn333) and the cardoon ( Cynara cardunculus L.) aspartic proteinase, cardosin A (Asn70 and Asn363). The oligosaccharides from each site were released from peptide pools by enzymatic hydrolysis with peptide‐N‐glycanase A or by hydrazinolysis and their structures were determined by exoglycosidase sequencing combined with matrix‐assisted laser desorption/ionization time of flight mass spectrometry. It was observed that 6% of the oligosaccharides from the first glycosylation site of cardosin A are of the oligomannose type. Modified type glycans with proximal Fuc and without Xyl account for about 82%, 14% and 3% of the total oligosaccharides from the first and the second glycosylation sites of cardosin A and from H. vulgare aspartic proteinase, respectively. Oligosaccharides with Xyl but without proximal Fuc were only detected in the latter proteinase (4%). Glycans with proximal Fuc and Xyl account for 6%, 86% and 92% of the total oligosaccharides from the first and second glycosylation sites of cardosin A and from H. vulgure aspartic proteinase, respectively.