Glycoproteomic survey of Mammillaria gracillis tissues grown in vitro.
The structure elucidation of protein-linked N-glycans in plants has raised interest in the past years due to remarkable physiological roles attributed to these modifications. However, little information about the glycoprotein patterns related to plant cell differentiation, dedifferentiation and transformation is available. In this work, the use of two-dimensional polyacrylamide gel electrophoresis in conjunction with matrix assisted laser/desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) for the characterization of carbohydrates released from plant glycoproteins is described. Proteins from different Mammillaria tissues (shoot, callus, hyperhydric regenerant, and TW tumor) were separated by 2D SDS-polyacrylamide gel electrophoresis, transferred to a nitrocellulose membrane and incubated with Con A to detect N-glycosylated proteins. To discover if the same protein can have various N-glycan structures depending on the organization status of the tissue, the selected glycoprotein spot, which was common for all investigated tissues, was excised from the gels and digested by PNGase A. The released oligosaccharides were analyzed by MALDI-TOF-MS. The results obtained in this study indicate that the N-glycosylation pattern of the protein is clearly dependent on level of plant tissue organization and can be related to the specific morphogenic status.
Balen
Biljana
B
Department of Molecular Biology, Faculty of Science, University of Zagreb, Horvatovac 102a, 10000 Zagreb, Croatia. bbalen@zg.biol.pmf.hr
Krsnik-Rasol
Marijana
M
Zamfir
Alina D
AD
Milosević
Jadranka
J
Vakhrushev
Sergey Y
SY
Peter-Katalinić
Jasna
J
D029421:Cactaceae / Q000737:chemistry* / Q000166:cytology* / Q000502:physiology; D002478:Cells, Cultured; D006023:Glycoproteins / Q000032:analysis*; D066298:In Vitro Techniques; D009369:Neoplasms / Q000737:chemistry / Q000473:pathology; D010449:Peptide Mapping; D010940:Plant Proteins / Q000032:analysis*; D018520:Plant Shoots / Q000737:chemistry / Q000166:cytology; D040901:Proteomics; D012038:Regeneration
D006023:Glycoproteins; D010940:Plant Proteins