title: Purification of a secreted lectin from Andrias davidianus skin and its antibacterial activity.
abstract: A lectin secreted from Andrias davidianus skin (ADL) was purified by affinity chromatography on porcine stomach mucin (type III) (PSM)-crosslinked albumin, followed by gel filtration on Sephadex G-100 and HPLC on TSK gel G3000PWXL. The purified lectin was found to be a dimeric protein, as revealed by SDS-PAGE and MALDI-TOF analysis. SDS-PAGE showed that the ADL protein had a molecular mass of 17 kDa. ADL produced an 8.5 kDa band when examined using SDS-PAGE under reducing conditions. ADL agglutinated native and trypsinized human B erythrocytes. The hemagglutination activity was inhibited by glycoproteins, such as PSM and asialo-PSM, but not by any of the monosaccharides tested. The activity was stable between 4 °C and 50 °C. Significant ADL activity was observed between pH 4–5. The lectin reaction did not depend on the presence of the divalent cation Ca2+ or Mg2+. The N-terminal ADL sequence was determined to be VGYTVGATPM. The lectin exhibited antibacterial activity, involving growth and respiration inhibition in Escherichia coli, Enterobacter aerogenes, Staphylococcus aureus, Bacillus subtilis and Shewanella sp. Furthermore, ADL showed inhibition activity against the yeast Saccharomyces cerevisiae. These findings suggest that ADL plays an important role in the innate immunity of A. davidianus on the body surface.
authors.affiliation: College of Food Science and Engineering, Dalian Ocean University, Dalian 116023, PR China.
authors.forename: Min
authors.initials: M
authors.lastname: Qu
authors.forename: Changqing
authors.initials: C
authors.lastname: Tong
authors.forename: Liang
authors.initials: L
authors.lastname: Kong
authors.forename: Xin
authors.initials: X
authors.lastname: Yan
authors.forename: Oleg V
authors.initials: OV
authors.lastname: Chernikov
authors.forename: Pavel A
authors.initials: PA
authors.lastname: Lukyanov
authors.forename: Qiao
authors.initials: Q
authors.lastname: Jin
authors.forename: Wei
authors.initials: W
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000494:pharmacology*; D000935:Antifungal Agents / Q000737:chemistry / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects; D050260:Carbohydrate Metabolism; D037102:Lectins / Q000737:chemistry* / Q000145:classification / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000187:drug effects; D012867:Skin / Q000378:metabolism*; D014562:Caudata / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D000935:Antifungal Agents; D037102:Lectins