Processing of the 2S storage protein pronapin in Brassica napus and in transformed tobacco.
The 2S protein napin in Brassica napus is synthesized as a proprotein from which an N-terminal an an internal propeptide are removed. In order to investigate the mechanism of 2S storage-protein processing, N-terminal sequences were determined for the light and heavy chains of all major napin isoforms isolated. Mutants of a napin gene, with deletions of different portions of the propeptides, were transformed into tobacco and napin protein was isolated. Napin light and heavy chains were separated and their N-terminal amino acid sequences determined. Further, the C-terminal residues of one napin isoform isolated from B. napus and one mutant napin isolated from tobacco were deduced from molecular-mass determinations of the constituents chains. Analyses suggested that the two propeptides are exposed at the surface of the proprotein. The light chain is processed to the correct length independent of the amino acid sequence in the N-terminal propeptide and the processing site. The internal propeptide is attacked by endoproteases. Aminopeptidases and carboxypeptidases then digest portions of the propeptide to the extent allowed by the primary and the three-dimensional structures, often resulting in 2S protein chains with partly frayed ends.
Murén
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Department of Cell Research, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Ek
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Rask
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D055516:2S Albumins, Plant; D000595:Amino Acid Sequence; D001483:Base Sequence; D001937:Brassica / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D010947:Plants, Toxic; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D012639:Seeds / Q000378:metabolism; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D014026:Nicotiana / Q000235:genetics; D014170:Transformation, Genetic
D055516:2S Albumins, Plant; D010940:Plant Proteins; D011498:Protein Precursors; C050363:napin protein, Brassica napus