Apoptosis-inducing effect and structural basis of Polygonatum cyrtonema lectin and chemical modification properties on its mannose-binding sites.
Polygonatum cyrtonema Lectin (PCL), which is classified as a monocot mannose-binding lectin, has received great regards for its uniquely biological activities and potentially medical applications in cancer cells. This paper was initially aimed to study apoptosis of PCL on Hela cells. Thus, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) method was carried out. Through observation of cell morphologic changes and Lactate dehydrogenase (LDH) activity-based cytotoxicity assays, PCL induced HeLa cell apoptosis in a dose-dependent manner. To further gain structural basis, multiple alignments, homology modeling and docking experiments were performed to analyze the correlation between its biological activities and mannose-binding sites. Eventually, considering docking data, chemical modification properties on the three mannose-binding sites were analyzed by a series of biological experiments (e.g., hemagglutinating and mitogenic activity assays, fluorescence and Circular Dichrosim (CD) spectroscopy) to profoundly identify the role of some key amino acids in the structure-function relationship of PCL.
Liu
Bo
B
College of Life Sciences, Sichuan University, Chengdu, China. jinkubao@yahoo.com
Xu
Xiao-Chao
XC
Cheng
Yan
Y
Huang
Jian
J
Liu
Yan-Hong
YH
Liu
Zhen
Z
Min
Ming-Wei
MW
Bian
He-Jiao
HJ
Chen
Jing
J
Bao
Jin-Ku
JK
D000595:Amino Acid Sequence; D017209:Apoptosis / Q000187:drug effects*; D001665:Binding Sites; D002942:Circular Dichroism; D006367:HeLa Cells; D006801:Humans; D006836:Hydro-Lyases / Q000378:metabolism; D037601:Mannose-Binding Lectin / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D037121:Plant Lectins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D031384:Polygonatum / Q000737:chemistry*; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship
D037601:Mannose-Binding Lectin; D037121:Plant Lectins; D006836:Hydro-Lyases; C045581:lactate dehydratase