Anti-HIV I/II activity and molecular cloning of a novel mannose/sialic acid-binding lectin from rhizome of Polygonatum cyrtonema Hua.
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.
College of Life Science, Sichuan University, Chengdu 610064, China.
Jie
J
An
Jin-Zhi
JZ
Liu
Chuan-Fang
CF
Wu
Jian
J
Li
Lei
L
Dai
Els
E
Van Damme
Jan
J
Balzarini
Erik
E
De Clercq
Fang
F
Chen
Jin-Ku
JK
Bao
D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000494:pharmacology*; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis; D015497:HIV-1 / Q000187:drug effects* / Q000378:metabolism; D015498:HIV-2 / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D037102:Lectins / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D037601:Mannose-Binding Lectin / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D037241:Mannose-Binding Lectins / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D037121:Plant Lectins / Q000737:chemistry; D027343:Rhizome / Q000737:chemistry*; D063265:Sialic Acid Binding Immunoglobulin-like Lectins
D000998:Antiviral Agents; D018076:DNA, Complementary; D037102:Lectins; D037601:Mannose-Binding Lectin; D037241:Mannose-Binding Lectins; D037121:Plant Lectins; D063265:Sialic Acid Binding Immunoglobulin-like Lectins; C101306:lectin-related protein, Polygonatum