Biochemical characterization and cellular localization of 11S type storage proteins in olive (Olea europaea L.) seeds.
The composition of seed storage proteins (SSPs) in olive endosperm and cotyledon has been analyzed. Precursor forms of these proteins are made up of individual proteins, which have been purified to homogeneity and further named p1-p5 (20.5, 21.5, 25.5, 27.5, and 30 kDa, respectively). N-terminal sequences of p1 and p2 proteins displayed relevant homology to the basic subunit of the 11S family of plant SSPs (legumins). Two-dimensional polyacrylamide gel electrophoresis experiments allowed us to verify the basic character of p1 and p2 and the acidic character of p3, p4, and p5 proteins. In addition, the putative presence of highly similar isoforms or posttranslational modifications of these polypeptides was detected. As a result, a model describing the putative association of p1-p5 proteins into subunits of alpha(acidic)/beta(basic) type has been proposed. Solubility experiments have shown that the majority of these olive seed proteins from the 11S storage protein family are extracted with aqueous alcohol and only partially with water and diluted saline solutions, therefore suggesting their similarity to prolamines. Moreover, no visible differences were found in either subunit composition or 11S proteins mass among six olive cultivars examined. This result suggests that the synthesis of storage proteins is highly conserved in this plant species. By using a rabbit antiserum raised to p1 protein, the proteins have also been immunolocalized in olive seed tissues, showing that they accumulate in conspicuous protein bodies present in both the endosperm and the cotyledon.
de Dios Alché
Juan
J
Departamento de Bioquímica, Biología Celular y Molecular de Plantas, Estación Experimental del Zaidín, CSIC, Profesor Albareda 1, E-18008 Granada, Spain.
Jiménez-López
José C
JC
Wang
Wei
W
Castro-López
Antonio J
AJ
Rodríguez-García
Maria I
MI
D000595:Amino Acid Sequence; D000918:Antibody Specificity; D004591:Electrophoresis, Polyacrylamide Gel; D007106:Immune Sera; D015151:Immunoblotting; D008854:Microscopy, Electron; D031658:Olea / Q000737:chemistry*; D010940:Plant Proteins / Q000032:analysis* / Q000737:chemistry* / Q000276:immunology; D012639:Seeds / Q000737:chemistry* / Q000648:ultrastructure; D012995:Solubility
D007106:Immune Sera; D010940:Plant Proteins