Anti-HIV Ⅰ/Ⅱ Activity and Molecular Cloning of a Novel Mannose/Sialic Acidbinding Lectin from Rhizome of Polygonatum cyrtonema Hua  

作　　者：Jie AN Jin-Zhi LIU Chuan-Fang WU Jian LI Lei DAI Els Van DAMME Jan BALZARINI Erik De CLERCQ Fang CHEN Jin-Ku BAO 

机构地区：[1]College of Life Science, Sichuan University, Chengdu 610064, China [2]Department of Molecular Biotechnology, Ghent University, 9000 Gent, Belgium [3]Rega Institute for Medical Research, Katholieke University Leuven, B-3000 Leuven, Belgium

出　　处：《Acta Biochimica et Biophysica Sinica》2006年第2期70-78,共9页生物化学与生物物理学报（英文版）

基　　金：This work was supported by a grant from the National Natural Science Foundation of China （No. 30000032）

摘　　要：The anti-human immunodeficiency virus （HIV） Ⅰ/Ⅱ activity of a mannose and sialic acid binding lectin isolated from rhizomes of Polygonatum cyrtonema Hua was elucidated by comparing its HIV infection inhibitory activity in MT-4 and CEM cells with that of other mannose-binding lectins （MBLs）. The anti-HIV activity of Polygonatum cyrtonema Hua lectin （PCL） was 10- to 100-fold more potent than other tested MBLs, but without significant cytotoxicity towards MT-4 or CEM cells. To amplify cDNA of PCL by 3＇/5＇-rapid amplification of cDNA ends （RACE）, the 30 amino acids of N-terminal were determined by sequencing and the degenerate oligonucleotide primers were designed. The full-length cDNA of PCL contained 693 bp with an open reading frame encoding a precursor protein of 160 amino acid residues, consisting of a 28-residue signal peptide, a 22-residue C-terminal cleavage peptide and a 110-residue mature polypeptide which contained three tandemly arranged subdomains with an obvious sequence homology to the monocot MBL. However, only one active mannose-binding site （QDNVY） was found in subdomain Ⅰ of PCL, that of subdomain Ⅱ and Ⅲ changed to HNNVY and PDNVY, respectively. There was no intron in PCL, which was in good agreement with other monocot MBLs. Molecular modeling of PCL indicated that its three-dimensional structure resembles that of the snowdrop agglutinin. By docking, an active sialic acid-binding site was found in PCL. The instabilization of translation initiation region （TIR） in mRNA of PCL benefits its high expression in rhizomes.The anti-human immunodeficiency virus （HIV） Ⅰ/Ⅱ activity of a mannose and sialic acid binding lectin isolated from rhizomes of Polygonatum cyrtonema Hua was elucidated by comparing its HIV infection inhibitory activity in MT-4 and CEM cells with that of other mannose-binding lectins （MBLs）. The anti-HIV activity of Polygonatum cyrtonema Hua lectin （PCL） was 10- to 100-fold more potent than other tested MBLs, but without significant cytotoxicity towards MT-4 or CEM cells. To amplify cDNA of PCL by 3＇/5＇-rapid amplification of cDNA ends （RACE）, the 30 amino acids of N-terminal were determined by sequencing and the degenerate oligonucleotide primers were designed. The full-length cDNA of PCL contained 693 bp with an open reading frame encoding a precursor protein of 160 amino acid residues, consisting of a 28-residue signal peptide, a 22-residue C-terminal cleavage peptide and a 110-residue mature polypeptide which contained three tandemly arranged subdomains with an obvious sequence homology to the monocot MBL. However, only one active mannose-binding site （QDNVY） was found in subdomain Ⅰ of PCL, that of subdomain Ⅱ and Ⅲ changed to HNNVY and PDNVY, respectively. There was no intron in PCL, which was in good agreement with other monocot MBLs. Molecular modeling of PCL indicated that its three-dimensional structure resembles that of the snowdrop agglutinin. By docking, an active sialic acid-binding site was found in PCL. The instabilization of translation initiation region （TIR） in mRNA of PCL benefits its high expression in rhizomes.

关 键 词：Polygonatum cyrtonema HUA mannose-binding lectin anti-human immunodeficiency virus（HIV） Ⅰ/Ⅱ molecular cloning 3＇/5＇-rapid amplification of cDNA ends （RACE） sequence alignment molecular modeling docking 

分 类 号：R512.91[医药卫生—内科学]