Open AccessThree Glycosylated Polypeptides Secreted by Several Embryogenic Cell Cultures of Pine Show Highly Specific Serological Affinity to Antibodies Directed against the Wheat Germin Apoprotein Monomer
Author(s) -
Jean-Marc Domon,
Bernard Dumas,
Éric Lainé,
Yves Meyer,
Alain David,
Hélène N. David
Publication year - 1995
Publication title -
plant physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.554
H-Index - 312
eISSN - 1532-2548
pISSN - 0032-0889
DOI - 10.1104/pp.108.1.141
Subject(s) - antiserum , biochemistry , polyclonal antibodies , electroblotting , glycoprotein , biology , complementary dna , peptide sequence , amino acid , extracellular , gel electrophoresis , microbiology and biotechnology , antibody , polyacrylamide gel electrophoresis , gene , enzyme , immunology
Embryogenic tissues of Pinus caribaea Morelet var hondurensis produce extracellular proteins; among them germins have been identified. Two-dimensional electrophoresis followed by electroblotting onto a polyvinylidene difluoride membrane allowed isolation and N-terminal amino acid sequencing of extracellular GP111, which is present within the five embryogenic cell lines studied. The amino acid sequence showed strong homologies with the sequences of germins deduced from cDNA sequencing, starting at the same amino acid position but one, compared with other sequences of mature germins deduced from protein sequencing. Immunoblots of embryogenic and nonembryogenic extracellular proteins indicated that the polypeptide GP111 plus two others with similar relative molecular mass values are present in embryogenic cell lines but not in nonembryogenic ones. They were recognized by an antiserum raised against the nonglycosylated monomer of wheat germin. The cross-reaction between pine and wheat apoproteins was highly specific. An antiserum against the glycosylated pentameric germin-like protein (an oxalate oxidase) of barley cross-reacted with all three, as well as with several other glycosylated polypeptides.