title: The first trimeric Galanthus nivalis agglutinin-related lectin of Orchidaceae was found in Dendrobium pendulum: purification, characterization, and effects of stress factors.
abstract: Trimeric Galanthus nivalis agglutinin-related lectin of Orchidaceae with two conformational forms was first studied in Dendrobium pendulum . It was highly expressed by stress factors. Using mannan-agarose column chromatography, a mannose-binding protein was purified from Dendrobium pendulum Roxb. pseudobulb. After heating in the presence of sodium dodecyl sulfate (SDS) with or without 2-mercaptoethanol, the protein showed one band with molecular mass of 14.0 kDa on SDS-polyacrylamide gel electrophoresis (PAGE). Without heating, three bands were found at positions of 14.0, 39.4, and 41.5 kDa, but a higher amount of 39.4 and 41.5 kDa protein bands were seen in the presence of 2-mercaptoethanol. Liquid chromatography-tandem mass spectrometry and database search indicated that the 14.0 kDa protein band contained three peptide fragments identical to parts of a lectin precursor from Dendrobiu m findleyanum Parish & Rchb.f. Native-PAGE and Ferguson plot showed that the purified protein had two native forms with molecular masses of 44.2 and 45.3 kDa, indicating three 14.0 kDa polypeptide subunits. The purified protein exhibited the agglutination activity with trypsinized chicken erythrocytes. It was then recognized as a Galanthus nivalis agglutinin-related lectin and named D. pendulum agglutinin (DPA). Using reverse transcription-polymerase chain reaction and DNA sequencing, the deduced amino acid sequence of DPA precursor showed the highest homology (96.4%) with a lectin precursor of D. findleyanum and contained three mannose-binding sites. Greater amounts of DPA were found when the pseudobulbs were treated with stress factors including ultraviolet light, abscisic acid, hydrogen peroxide, and acetylene gas.
authors.affiliation: Department of Biochemistry, Faculty of Science, Khon Kaen University, Muang, Khon Kaen, 40002, Thailand.
authors.forename: Patthraporn
authors.initials: P
authors.lastname: Siripipatthana
authors.forename: Narumon
authors.initials: N
authors.lastname: Phaonakrop
authors.forename: Sittiruk
authors.initials: S
authors.lastname: Roytrakul
authors.forename: Gulsiri
authors.initials: G
authors.lastname: Senawong
authors.forename: Rasika G
authors.initials: RG
authors.lastname: Mudalige-Jayawickrama
authors.forename: Nison
authors.initials: N
authors.lastname: Sattayasai
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D031667:Dendrobium / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D037601:Mannose-Binding Lectin / Q000378:metabolism; D037241:Mannose-Binding Lectins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D008623:Mercaptoethanol / Q000494:pharmacology; D008969:Molecular Sequence Data; D037121:Plant Lectins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D055503:Protein Multimerization / Q000187:drug effects; D012015:Reference Standards; D017386:Sequence Homology, Amino Acid; D013312:Stress, Physiological / Q000187:drug effects
chemical_list: D018076:DNA, Complementary; D037102:Lectins; D037601:Mannose-Binding Lectin; D037241:Mannose-Binding Lectins; D037121:Plant Lectins; C063117:snowdrop lectin; D008623:Mercaptoethanol