1. Characterization and cDNA Cloning of Monomeric Lectins That Correspond to the B-Chain of a Type 2 Ribosome-Inactivating Protein from the Bark of Japanese Elderberry (Sambucus sieboldiana)
- Author
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Misa Yato, Shigeru Hisajima, Naoto Shibuya, Maria Angeles Rojo, Naoko Ishii-Minami, Takeshi Yamaguchi, Hanae Kaku, and Eiichi Minami
- Subjects
Erythrocyte Aggregation ,Glycosylation ,Ribosome Inactivating Proteins ,Biochemistry ,Homology (biology) ,Cloning, Molecular ,Fetuins ,N-Glycosyl Hydrolases ,Plant Proteins ,chemistry.chemical_classification ,biology ,Sambucus sieboldiana ,Serum Albumin, Bovine ,General Medicine ,Chromatography, Ion Exchange ,Amino acid ,Ribosome Inactivating Proteins, Type 2 ,Blotting, Southern ,Sambucus ,Chromatography, Gel ,Plant Bark ,Electrophoresis, Polyacrylamide Gel ,Rabbits ,alpha-Fetoproteins ,Plant Lectins ,Protein Binding ,DNA, Complementary ,Molecular Sequence Data ,Carbohydrates ,Asialoglycoproteins ,Enzyme-Linked Immunosorbent Assay ,stomatognathic system ,Complementary DNA ,Animals ,Amino Acid Sequence ,Molecular Biology ,Sequence Homology, Amino Acid ,cDNA library ,Ribosome-inactivating protein ,Galactose ,Lectin ,Sequence Analysis, DNA ,biology.organism_classification ,Molecular biology ,Molecular Weight ,stomatognathic diseases ,Isoelectric point ,chemistry ,Protein Biosynthesis ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,biology.protein - Abstract
Two monomeric lectins, SSA-b-3 and SSA-b-4, were purified from the bark tissue of Japanese elderberry, Sambucus sieboldiana. SDS-PAGE of the purified lectins showed the presence of single bands of 35 and 33 kDa for SSA-b-3 and SSA-b-4, respectively, irrespective of the presence of reducing agent. MS analysis as well as gel filtration of these lectins indicated that they exist mostly as monomeric lectins. Analysis of the N-terminal amino acid sequences of SSA-b-3 and SSA-b-4 yielded an identical sequence, indicating their close structural relationship. Four cDNA clones with extensive homology were obtained from the bark cDNA library and indicated to encode SSA-b-3 or SSA-b-4 from the comparison with the N-terminal sequences of these lectins. These clones were classified into two groups, three for SSA-b-3 and one for SSA-b-4, based on the predicted isoelectric points. The amino acid sequences of the encoded polypeptides were almost identical with the B-chain of a type 2 ribosome-inactivating protein from the same bark tissue, sieboldin-b, except for the absence of a small peptide containing a cystein residue, which is critical for the heteromeric dimerization with an A-subunit. Carbohydrate binding specificity and biological activity of these lectins are also reported.
- Published
- 2004
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