Pea legumin overexpressed in wheat endosperm assembles into an ordered paracrystalline matrix.
Legumin, a major component of pea seed storage vacuoles, is synthesized by a number of paralogous genes. The polypeptides are cleaved posttranslationally and can form mixed hexamers. This heterogeneity hampers structural studies, based on the production of hexamer crystals in vitro. To study a single type of homogenous legumin we produced pea legumin A in transgenic wheat (Triticum aestivum) endosperm where prolamins are predominant and only small amounts of globulins accumulate in separate inclusions. We demonstrated that the legumin precursor was cleaved posttranslationally and we confirmed assembly into 11S hexamers. Legumin was deposited within specific regions of the inclusion bodies. Angular legumin crystals extended from the inclusion bodies into the vacuole, correlating with the high legumin content. This suggests that the high-level production of a single type of legumin polypeptide resulted in the spontaneous formation of crystals in vivo. The use of a heterologous cereal system such as wheat endosperm to produce, isolate, and recrystallize homogenous 11S legume globulins offers exciting possibilities for structural analysis and characterization of these important seed storage proteins.
Stöger
E
E
Molecular Biotechnology Unit, John Innes Centre, Norwich Research Park, Colney Lane, Norwich NR4 7UH, United Kingdom.
Parker
M
M
Christou
P
P
Casey
R
R
D018076:DNA, Complementary; D017930:Genes, Reporter; D005966:Glucuronidase / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D005983:Glutens / Q000031:analogs & derivatives* / Q000235:genetics; D007150:Immunohistochemistry; D007438:Introns; D016253:Microscopy, Immunoelectron; D018532:Pisum sativum / Q000235:genetics*; D010940:Plant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000378:metabolism; D012639:Seeds / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D014908:Triticum / Q000166:cytology / Q000235:genetics* / Q000378:metabolism*; D014452:Ubiquitins / Q000235:genetics; D000097793:Legumins
D018076:DNA, Complementary; D005966:Glucuronidase; D005983:Glutens; D010940:Plant Proteins; D014452:Ubiquitins; D000097793:Legumins; C037302:glutenin