Subcellular localization of the 2S globulin narbonin in seeds of Vicia narbonensis.
Narbonin is a 2S protein from the globulin fraction of narbon bean (Vicia narbonensis L.) cotyledons. Its amino acid composition and the pattern of its regulated accumulation in developing seeds led to the suggestion that narbonin could be a storage protein. Therefore, it was expected to be present in protein bodies of the storage tissue cells. Comparison of the cDNA-derived amino acid sequence with a directly determined partial N-terminal sequence revealed that the primary translation product of narbonin mRNA lacks a transient N-terminal signal peptide (V.H. Nong et al., 1995, Plant Mol Biol 28: 61 - 72). Narbonin polypeptides that had been synthesized in a cell-free translation system supplemented with dog pancreas microsomes were not protected against degradation by posttranslationally added proteases (protease protection assay). In accordance with the lack of a signal peptide this indicates that the polypeptide was not cotranslationally sequestered into the microsomes. The protein-body fraction that had been isolated from mature narbon bean cotyledons by a non-aqueous gradient centrifugation procedure was free of narbonin; this was found in the soluble cell fraction. In electron micrographs, narbonin could be localized in the cytoplasm using the immuno gold-labelling technique. Previously, it had already been shown that narbonin is too slowly degraded during narbon bean germination to act as a storage protein. From all these results it has to be concluded that narbonin is a cytoplasmic protein which does not belong to the storage proteins in the restricted sense. Other possible functions are discussed.
Steffens
P
P
Institut für Pflanzengenetik und Kulterpflanzenforschung, Gatersleben, Germany.
Van
H N
HN
Hillmer
S
S
Saalbach
I
I
Müntz
K
K
D000818:Animals; D002474:Cell-Free System; D018076:DNA, Complementary; D004285:Dogs; D007887:Fabaceae / Q000378:metabolism / Q000648:ultrastructure*; D005916:Globulins / Q000032:analysis* / Q000096:biosynthesis* / Q000737:chemistry; D016253:Microscopy, Immunoelectron; D008861:Microsomes / Q000378:metabolism; D010179:Pancreas / Q000378:metabolism; D014674:Plant Proteins, Dietary / Q000032:analysis* / Q000096:biosynthesis* / Q000737:chemistry; D010946:Plants, Medicinal; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000378:metabolism; D012639:Seeds / Q000737:chemistry / Q000648:ultrastructure; D013347:Subcellular Fractions / Q000378:metabolism / Q000648:ultrastructure
D018076:DNA, Complementary; D005916:Globulins; D014674:Plant Proteins, Dietary; D012333:RNA, Messenger; C066193:narbonin