Journal Article

The first trimeric Galanthus nivalis agglutinin-related lectin of Orchidaceae was found in Dendrobium pendulum: purification, characterization, and effects of stress factors

Sattayasai, Nison, Senawong, Gulsiri, Siripipatthana, Patthraporn, Roytrakul, Sittiruk, Phaonakrop, Narumon, Mudalige-Jayawickrama, Rasika G.

2015

Overview

Key message 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.

• Publisher: Springer Berlin Heidelberg,Springer Nature B.V
• Subject: abscisic acid / acetylene / agglutination / agglutinins / Amino Acid Sequence / amino acid sequences / Amino acids / Base Sequence / beta-mercaptoethanol / Biomedical and Life Sciences / Biotechnology / Cell Biology / chickens / Dendrobium / Dendrobium - chemistry / DNA, Complementary - genetics / Electrophoresis, Polyacrylamide Gel / erythrocytes / Galanthus nivalis / heat / Hot Temperature / Hydrogen peroxide / lectins / Lectins - chemistry / Lectins - isolation & purification / Lectins - metabolism / Life Sciences / Liquid chromatography / Mannose-Binding Lectin - metabolism / Mannose-Binding Lectins - chemistry / Mannose-Binding Lectins - isolation & purification / Mannose-Binding Lectins - metabolism / Mass spectrometry / Mercaptoethanol - pharmacology / Molecular Sequence Data / molecular weight / Original Paper / Plant Biochemistry / Plant Lectins - chemistry / Plant Lectins - isolation & purification / Plant Lectins - metabolism / Plant Sciences / polyacrylamide gel electrophoresis / polypeptides / Protein Multimerization - drug effects / Reference Standards / reverse transcriptase polymerase chain reaction / sequence analysis / Sequence Homology, Amino Acid / sodium dodecyl sulfate / Stress, Physiological - drug effects / tandem mass spectrometry / Ultraviolet radiation