caa a22 4500 477101259 CHVBK 20180405111542.0 cr unu---uuuuu 170330e19960201xx s 000 0 eng 10.1007/BF00206247 doi (NATIONALLICENCE)springer-10.1007/BF00206247 Characterisation of a developmentally related polypeptide with glutelin solubility characteristics from Lupinus albus L [Elektronische Daten] [Júlia Costa, David Ashford, Cândido Pinto Ricardo] Proteins from Lupinus albus L. cv. Rio Maior seeds were fractionated according to solubility criteria. Patterns of concanavalin A (ConA)-binding polypeptides from the different classes, albumins, globulins, glutelins and prolamins, were established by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Two bands of apparent molecular masses of 29 and 23.5 kDa with glutelin solubility characteristics bound the lectin. The 23.5-kDa band was separated by two-dimensional electrophoresis into two components: one glycosylated and heterogeneous with an isoelectric point of approx. 10 (designated as G23) and another, not detected with ConA, precipitating in the first dimension. The amino acid and hexosamine analysis of G23 showed that it is particularly rich in Gly (11.2%), Glx (10.0%), Ser (9.0%), Leu (8.2%), Asx (7.5%), and Pro (6.7%) and that it has a considerable content of the sulphur-containing amino acids Met (2.0%) and Cys (5.8%) and contains glucosamine. The determined N-terminal amino acid sequence of G23 was: 1KG(R)V5KGTGD10(T)PXXV15XLY(N)R20T, and this had no significant similarity to any of the amino acid sequences contained in the data bank SWISS-PROT 26. The glycoprotein G23 was completely deglycosylated with peptide-N-glycosidase F, yielding a homogeneous 21-kDa polypeptide composed of approximately 191 amino acids. The structures of the major N-linked neutral oligosaccharides of G23, determined by exoglycosidase sequencing, were as follows: Manα2Manα6(Manα3) Manα6(Manα2Manα2Manα3)Manβ4GlcNAcβ4GlcNAc (13%); ± Manα2Manα6(Manα3)Manα6(± Manα2 Manα2 Manα3)Manβ4GlcNAcβ4GlcNAc (29%); Manα6(Manα3) Manα6(Manα2Manα3)Manβ4GlcNAcβ4GlcNAc (13%); Manα6(Manα3)Manα6(Manα3)Manβ4GlcNAcβ4GlcNAc (16%); Manα6(Manα3)(Xylβ2)Manβ4GlcNAc β4GlcNAc (28%). Changes in G23 abundance during seed development, germination and seedling growth were monitored with a specific antibody. The glycoprotein G23 started to accumulate appreciably during seed formation between the 40th and the 50th days after anthesis and was detected following seed imbibition, until the 9th day in cotyledons, the 2nd day in roots and the 4th day in hypocotyls and leaves. Springer-Verlag, 1996 Germination nationallicence Glutelin nationallicence Glycoprotein nationallicence Leguminosae nationallicence Lupinus nationallicence Seed development nationallicence ConA : concanavalin A nationallicence Endo H : endo-N-acetyl-β-d-glucosaminidase H nationallicence GlcNAc : N-acetylglucosamine nationallicence gu : glucose unit nationallicence IEF : isoelectric focusing nationallicence Man : mannose nationallicence NEPHGE : non-equilibrium pH gradient electrophoresis nationallicence PNGase F : peptide-N-glycosidase F nationallicence PVDF : polyvinylidenedifluoride nationallicence Xyl : xylose nationallicence Costa Júlia Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Rua da Quinta Grande, 6, Apart. 127, 2780, Oeiras, Portugal aut Ashford David Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, South Parks Road, OX1 3QU, Oxford, UK aut Pinto Ricardo Cândido Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Rua da Quinta Grande, 6, Apart. 127, 2780, Oeiras, Portugal aut Planta Springer-Verlag 198/2(1996-02-01), 221-229 0032-0935 198:2<221 1996 198 425 https://doi.org/10.1007/BF00206247 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00206247 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Costa Júlia Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Rua da Quinta Grande, 6, Apart. 127, 2780, Oeiras, Portugal aut NATIONALLICENCE 700 1- Ashford David Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, South Parks Road, OX1 3QU, Oxford, UK aut NATIONALLICENCE 700 1- Pinto Ricardo Cândido Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Rua da Quinta Grande, 6, Apart. 127, 2780, Oeiras, Portugal aut NATIONALLICENCE 773 0- Planta Springer-Verlag 198/2(1996-02-01), 221-229 0032-0935 198:2<221 1996 198 425 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer