Glycosylation of proteins in endoplasmic reticulum during horseradish (Armoracia lapathifolia Gilib.) cellular differentiation in in vitro culture

Gospočić, Janko (2008) Glycosylation of proteins in endoplasmic reticulum during horseradish (Armoracia lapathifolia Gilib.) cellular differentiation in in vitro culture. Diploma thesis, Faculty of Science > Department of Biology.

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Abstract

In this work different horseradish transformed lines (tumour and teratoma) were compared with control samples of leaf tissue in electrophoretic pattern of total cell proteins and their glycosylation. Emphasis was put on the comparison of these tissues in structure and composition of N-glycans of endoplasmic reticulum proteins. Despite its central metabolic role, the endoplasmic reticulum is also a starting point of protein glycosylation in eukaryotic cells. Two forms of protein glycosylation are known, N- and Oglycosylation. N-glycosylation is a major modification of proteins in plant cells. Plant N-glycans can be classified into four basic groups: high-mannose-type, complex-type, paucimannosidic-type and hybrid-type. N-glycosylation is a well described process whereas O-glycosylatin is not well characterized. Glycoproteins were separated on SDS-PAGE and subsequently transferred onto nitrocellulose membrane. Glycosylated proteins were detected with Con A. N-glycans were further characterized by treating the membranes with different lectins (GNA, MAA, SNA, PNA, DSA, RCA I and UEA). The obtained results showed that glycosylation of total cell proteins and proteins of endoplasmic reticulum had been changed during horseradish cellular differentiation. Electron microscopy showed great differences in cell structure between leaf tissue, teratoma tissue (organized and unorganized) and tumour tissue. In all transformed horseradish lines plastids with irregular structure were noticed. The great similarity between tumour tissue and unorganized teratoma tissue was also observed. These two types of tissues had large abnormal nuclei and small plastids. In all types of tissues no differences in structure of endoplasmic reticulum were noticed.

Item Type: Thesis (Diploma thesis)
Keywords: horseradish, cellular differentiation, glycosylation, N-glycans, endoplasmic reticulum, lectins, electron microscopy
Supervisor: Balen, Biljana
Date: 2008
Number of Pages: 47
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Silvana Šehić
Date Deposited: 15 Sep 2014 09:43
Last Modified: 15 Sep 2014 09:43
URI: http://digre.pmf.unizg.hr/id/eprint/2757

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