The mode of CVN binding to oligosaccharides.

<p>An overview of the mode of CVN binding to oligosaccharides (<b>A</b>) No. 22 and (<b>B</b>) No. 28 is presented. The fuzzy blue blob indicates ligand exposure to the solvent. For oligosaccharide 22, which exhibited strong binding, Leu-1, Lys-3, Thy-7, Glu-23, Thr-25, Tyr-29 and Glu-101 were involved in the protein-ligand interaction. For oligosaccharide No. 28, which was characterized by weak binding, the docking simulation indicated that hydrogen bonds formed between the ligand and Leu-1, Gly-2, Lys-3, Thr-7, Thr-25 and Asn-93.</p

The anti-HIV-1 activity and MT-4 cytotoxicity of LCVNs (mean±SD, n = 3).

<p>IC₅₀, 50% viral inhibitory concentration;</p><p>CC₅₀, 50% cell inhibitory concentration;</p><p>SI, the ratio of CC₅₀ to IC₅₀.</p

Structural schematics of LCVN and the PEGylated product.

<p>The N-terminal glycine of LCVN and the serine-leucine joint between the linker and the CVN sequence are indicated. The blank rectangle represents the residual polypeptide of CVN. 10 KD mPEG-ALD was selectively reacted with the N-terminal α-amine of LCVN at different pKa values to create 10 K PEG-ALD-LCVN.</p

Binding specificities of CVN to PA-oligosaccharides.

<p>The optimum (<b>A</b>) pH and (<b>B</b>) reaction time for CVN binding to PA-heptasaccharide (No. 19, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0086455#pone-0086455-g001" target="_blank">Figure 1</a>) were determined via centrifugal ultrafiltration-HPLC, and subsequently the (<b>C</b>) binding activity (specificity) of CVN and the 24 oligosaccharides was measured. The right panel in (<b>C</b>) depicts the non-reducing terminal Manα₁₋₂Man moieties in the D1, D2 or D3 arms of the Man₇₋₉GlcNAc₂ glycans that participated in the binding. Two independent experiments were performed for each PA-oligosaccharide, and the binding activity is presented as the average of the duplicate assays.</p

MOE docking values for 53 oligosaccharides in the training set.

<p>x, the docking energy for 3GXY;</p><p>y, the docking energy for 2PYS.</p

The PA-oligosaccharide structures utilized for the molecular docking and experimental target binding assays with LCVNs.

<p>A panel of 53 oligosaccharides was selected to represent the diverse carbohydrate structures. All these oligosaccharides were utilized in the docking experiments. After the docking simulation, 24 oligosaccharides (indicated by asterisks) were selected for further analysis by the centrifugal ultrafiltration-HPLC assay.</p

Characterization of CVN binding to high mannose oligosaccharides.

<p>(<b>A</b>) The frequencies of the hot spot residues that target the 6 high mannose oligosaccharides were calculated. Leu-1 represents the N-terminal leucine in the B chain, which was involved in oligosaccharide binding with a frequency of 100%. (<b>B</b>) The total number of targeting residues (black column) and Manα₁₋₂Man binding residues (grey column) for CVN binding to the 6 oligosaccharides is presented. (<b>C</b>) The 3D model of CVN 3GXY binding to oligosaccharide No. 19 is illustrated. The 10 residues in CVN that were involved in the binding are colored in light pink, and their ligands are depicted in green. The hydrogen bonds are illustrated as dashed light pink lines.</p

LCVN fusion inhibitory activity.

<p>(<b>A</b>) Phase-contrast micrographs were obtained 24 h after co-culturing normal MOLT-4 and HIV-1-positive cells in the presence of LCVN. The HIV-1-induced multinucleated giant cells are indicated by black arrows. (<b>B</b>) The relative fusion inhibition rates (%) for LCVN and its derivatives were calculated (**<i>P</i><0.01 vs. CVN, * <i>P</i><0.05 vs. CVN).</p