Studies of the role of the propeptides of the Arabidopsis thaliana 2S albumin

• Published in: Plant physiology (Bethesda) 1993-06, Vol.102 (2), p.425-433
• Main Authors: D'Hondt, K, Damme, J. van, Bossche, C. van den, Leejeerajumnean, S, Rycke, R. de, Derksen, J, Vandekerckhove, J, Krebbers, E
• Format: Article
• Language: English
• Subjects: 2S Albumins, Plant; ALBUMINAS; ALBUMINE; Albumins; Albumins - genetics; Albumins - metabolism; Amino Acid Sequence; Amino acids; Antigens, Plant; Antiserum; Arabidopsis - genetics; Arabidopsis - metabolism; ARABIDOPSIS THALIANA; Biological and medical sciences; Cell Biology and Signal Transduction; Fundamental and applied biological sciences. Psychology; Gene Deletion; Genes, Plant; GRAINE; Immunohistochemistry; IMMUNOLOGIE; INMUNOLOGIA; Metabolism; Molecular Sequence Data; Nicotiana - genetics; Nitrogen metabolism; Plant cells; Plant physiology and development; Plant Proteins - genetics; Plant Proteins - metabolism; Plants; Plants, Genetically Modified; Plants, Toxic; Protein precursors; Protein Precursors - genetics; Protein Precursors - metabolism; Protein Processing, Post-Translational - genetics; Protein Sorting Signals - genetics; Protein Sorting Signals - metabolism; Proteins; PROTEOLISIS; PROTEOLYSE; Saccharomyces cerevisiae - genetics; Seeds; SEMILLA; Vacuoles; Yeasts
• ISSN: 0032-0889; 1532-2548
• DOI: 10.1104/pp.102.2.425

To investigate the possible roles of the Arabidopsis thaliana 2S albumin propeptides with respect to sorting, processing, and stability of the protein in plant cells, five gene constructions deleting or modifying the propeptides were made based on one of the genes encoding the Arabidopsis 2S albumin. These constructions were introduced into tobacco (Nicotiana tabacum) plants. Using subcellular fractionation and immunocytochemistry on ripe seeds, it was demonstrated that none of the propeptides was necessary for the sorting of the protein. Detailed protein-chemical analysis of the mature gene products indicated that, for all of the modified 2S albumin precursors made, the proteins were stably folded and correctly processed. However, the latter is less efficient when the internal fragment between the small and the large subunit is missing or when this internal fragment is changed. In an attempt to establish a rapid assay system for modified 2S albumin precursors, yeast cells were transformed with the same gene constructs. it was demonstrated that the processing machinery in yeast cells differs from that in plants, and, in a perhaps related observation, differences in stability of a particular modified protein were observed