title: Inhibitors of protein glycosylation or secretion change the pattern of extracellular proteins in suspension-cultured cells of Arabidopsis thaliana.
abstract: Cell walls are essential for plant development and morphogenesis. The majority of wall proteins are glycosylated, either as N- or O-glycans. Various inhibitors of glycosylation and secretion are used to determine the importance of wall proteins for the functioning of the walls. Tunicamycin is an inhibitor of the first enzyme in the N-glycosylation pathway, 3,4-dehydroproline inhibits peptidyl proline hydroxylation, and Brefeldin A is an inhibitor of vesicle trafficking, disrupting the delivery of wall polymers to the apoplast. In inhibitor-treated suspension-cultured Arabidopsis thaliana cells, qualitative and quantitative differences in the extracellular proteome were observed for both proteins secreted into medium or ionically-bound in the walls. Lack of O-glycosylation resulted in the selective protein loss from the extracellular compartments. Following tunicamycin treatment the secretion of additional proteins as well as ER-resident chaperones from the Hsp70 and Hsp90 families outside the protoplasts was noted. Moreover, changes in the proteolytic degradation pattern of culture filtrate proteins were also observed. Application of Brefeldin A resulted in transient and selective loss of individual proteins from the extracellular compartments of A. thaliana cell suspension. We conclude that post-translational modifications are vital for the proper functioning of wall proteins. N-glycosylation is crucial for their proper folding and stability. Extracellular compartments could also serve as a sink for improperly folded proteins during the unfolded protein response.
authors.affiliation: Department of Molecular and Cellular Biology, Adam Mickiewicz University, Miedzychodzka 5, 60-371 Poznań, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Luczak
authors.forename: Alina
authors.initials: A
authors.lastname: Bugajewska
authors.forename: Przemysław
authors.initials: P
authors.lastname: Wojtaszek
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D002473:Cell Wall / Q000378:metabolism*; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000378:metabolism*; D005110:Extracellular Space / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D006031:Glycosylation; D010940:Plant Proteins / Q000032:analysis / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D006023:Glycoproteins; D010940:Plant Proteins