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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Asialoglycoproteins / metabolism
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Blotting, Southern
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Carbohydrates / pharmacology
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Chromatography, Gel
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Chromatography, Ion Exchange
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Cloning, Molecular
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DNA, Complementary / chemistry
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DNA, Complementary / genetics
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Electrophoresis, Polyacrylamide Gel
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Enzyme-Linked Immunosorbent Assay
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Erythrocyte Aggregation / physiology
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Fetuins
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Galactose / metabolism
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Glycosylation
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Molecular Sequence Data
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Molecular Weight
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N-Glycosyl Hydrolases / genetics
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N-Glycosyl Hydrolases / metabolism
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N-Glycosyl Hydrolases / pharmacology
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Plant Bark / chemistry
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Plant Bark / genetics
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Plant Lectins / chemistry
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Plant Lectins / genetics*
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Plant Lectins / metabolism
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Plant Proteins / genetics
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Plant Proteins / metabolism
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Plant Proteins / pharmacology
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Protein Binding / drug effects
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Protein Biosynthesis / drug effects
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Rabbits
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Ribosome Inactivating Proteins
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Ribosome Inactivating Proteins, Type 2
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Sambucus / chemistry
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Sambucus / genetics*
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Sequence Analysis, DNA
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Sequence Homology, Amino Acid
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Serum Albumin, Bovine / metabolism
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
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alpha-Fetoproteins / metabolism
Substances
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Asialoglycoproteins
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Carbohydrates
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DNA, Complementary
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Fetuins
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Plant Lectins
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Plant Proteins
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Ribosome Inactivating Proteins, Type 2
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Sambucus nigra lectins
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alpha-Fetoproteins
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asialofetuin
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asialoglycophorin
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bovine serum albumin-galactose (39)
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sieboldin b
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Serum Albumin, Bovine
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N-Glycosyl Hydrolases
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Ribosome Inactivating Proteins
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Galactose
Associated data
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GENBANK/AB005718
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GENBANK/AB005719
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GENBANK/AB005720
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GENBANK/AB005721