title: Purification of a mannose-binding lectin Pinellia ternata agglutinin and its induction of apoptosis in Bel-7404 cells.
abstract: A novel high-throughput purification method for a monocot mannose-binding lectin, Pinellia ternata agglutinin (PTA), from tubers of P. ternata was established by mannose-Sephrose 4B affinity chromatography. The total protein was extracted from tubers of P. ternata using phosphate buffered saline (PBS) buffer. The extracted total protein was precipitated completely at 65% ammonium sulfate saturation and dissolved in different concentrations of NaCl solution to activate its binding affinity toward the column. PTA was bound to the affinity column by loading of the total protein into the column and elution using PBS buffer. The maximum purification yield (35.5mg/g) was obtained when PTA was treated with 25% (w/v) NaCl solution, and the purity of PTA analyzed by SDS-PAGE was ∼97%. The agglutination property of purified PTA was confirmed by mouse erythrocytes, which indicates its biological function. Nuclear staining assay and DNA fragmentation demonstrated that PTA could induce apoptosis of Bel-7404 cells, which further demonstrates its biological and pharmacological activities. Induction of apoptosis in the human tumor Bel-7404 cell line by PTA indicates its possible use in cancer therapy. The present investigation reports a significantly improved isolation method to obtain highly purified mannose-binding plant lectin proteins. The proposed method has great potential for industrial application because of its advantages, which include rapid isolation, high purity, high yield, low cost, and minimal requirement of chemical materials.
authors.affiliation: Institute of Bioengineering, College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhou
authors.forename: Yong
authors.initials: Y
authors.lastname: Gao
authors.forename: Shaowei
authors.initials: S
authors.lastname: Xu
authors.forename: Zhili
authors.initials: Z
authors.lastname: Yang
authors.forename: Tao
authors.initials: T
authors.lastname: Xu
keywords: Affinity purification; Apoptosis; High-throughput; Mannose; Pinellia ternata agglutinin