Proteolysis at the plasma membrane of tobacco roots: biochemical evidence and possible roles.
Plasma membrane-associated proteases (pm-proteases) exist principally in roots of Nicotiana tabacum cv. Samsun, whereas in plasma membrane (pm) vesicles prepared from leaves, protease activity was at the detection limit. Biochemical characterisation revealed a high diversity of particular hydrophobic pm-proteases indicating multiple functions in root tissue. One proportion of chromatographically separated proteases was split up by non-reducing SDS-PAGE in 8-12 single polypeptides, dependent on plant nitrogen nutrition. The active polypeptides could be grouped in those that were (i) inhibited, (ii) stimulated and (iii) independent of bivalent cations. Although, the total specific protease activity of various pm vesicles was almost identical, the composition and activity of individual polypeptides was dependent on nitrogen supply of the plants. Particularly, nitrogen deficiency stimulated the activity of high molecular mass proteases (125 kDa-97 kDa), whereas sufficient nitrate supply enhanced proteolytic activity of 90 kDa, 83 kDa and 65 kDa polypeptides. Endogenous proteolysis within pm vesicles suggested that at least partly protease substrates are localised within the same membrane. A comparison of polypeptides originated from proteolysis of pm vesicles and those exudated by roots into the external medium points to a role of root pm-proteases in the specific release of polypeptides into the rhizosphere.
Eick
Manuela
M
University Greifswald, Institute of Botany and Landscape Ecology, Grimmer Strasse 88, D-17487 Greifswald, Germany.
Stöhr
Christine
C
D002462:Cell Membrane / Q000378:metabolism*; D022162:Cytoplasmic Vesicles / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D008970:Molecular Weight; D009584:Nitrogen / Q000378:metabolism; D010455:Peptides / Q000378:metabolism*; D018515:Plant Leaves / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism*; D018517:Plant Roots / Q000378:metabolism*; D014026:Nicotiana / Q000378:metabolism*
D010455:Peptides; D010940:Plant Proteins; D009584:Nitrogen