31172315, 81271871) for J. Background The outer-tegument membrane covering the schistosome is definitely believed to preserve via the fusion of membranous vesicles. Fusion of biological membranes is definitely a fundamental process in all eukaryotic cells driven by formation of trans-SNARE (soluble N-ethylmaleimide-sensitive element attachment protein receptor) complexes through pairing of vesicle connected v-SNAREs (VAMP) with complementary t-SNAREs on target membranes. The purpose of this study was to characterize vesicle-associated membrane protein 2 (SjVAMP2) and to investigate its potential as a candidate vaccine against schistosomiasis. Strategy/Principal Findings The sequence of SjVAMP2 was analyzed, cloned, expressed and characterized. SjVAMP2 is definitely a member of the synaptobrevin superfamily harboring the v-SNARE coiled-coil homology website. RTCPCR analysis exposed that significantly higher SjVAMP2 levels were observed in 14-, 28- and 42-day-old worms, and SjVAMP2 expression was much higher in 42-day-old female worms than in those male worms. Additionally, the expression of SjVAMP2 was associated with membrane recovery in PZQ-treated worms. Immunostaining assay showed that SjVAMP2 was mainly distributed in the sub-tegument of the worms. Western blotting revealed that rSjVAMP2 showed strong immunogenicity. Purified rSjVAMP2 emulsified with ISA206 Docetaxel (Taxotere) adjuvant induced 41.5% and 27.3% reductions in worm burden, and 36.8% and 23.3% reductions in hepatic eggs in two indie trials. Besides, significantly higher rSjVAMP2-specific IgG, IgG1, IgG2a levels were detected in rSjVAMP2-vaccinated mice. Conclusion Our study indicated that SjVAMP2 is usually a potential vaccine candidate against and provided the basis for further investigations into the biological function of SjVAMP2. Introduction Schistosomiasis is an important parasitic disease epidemic in the tropics and subtropics that infects more than 200 million people in over 70 countries, causes an estimated 280,000 deaths annually, and endangers an additional 650 million people worldwide [1,2]. Moreover, schistosomiasis represents an increasing problem in non-endemic areas due to environmental change and the growing quantity of immigrants Rabbit polyclonal to ZBTB8OS and visitors [3,4]. (in our laboratory showed that vesicle-associated membrane protein 2 (VAMP2) is usually a tegument surface protein. Besides, VAMP2 is usually a key Docetaxel (Taxotere) part of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) that is required for membrane fusion. With few exceptions, the fusion of biological membranes, a fundamental process governing the transport of cargo molecules, such as the trafficked proteins, hormones, Docetaxel (Taxotere) and neurotransmitters throughout the secretory and endocytic pathways, is usually driven by the formation of trans-SNARE complexes [14,15]. SNAREs are compartment-specific proteins that consist of VAMP2 bound to the synaptic vesicle (v-SNARE), syntaxin and SNAP25 on the target membrane (t-SNARE). By pairing v-SNAREs with cognate t-SNAREs, a bundle of four helices (SNAREpins), three derived from the t-SNARE and the fourth from your cognate v-SNARE, is usually assembled, bringing the two lipid bilayers into close Docetaxel (Taxotere) proximity, finally culminating in membrane fusion [16,17]. SNAREpin assembly is usually a sequential, two-step folding pathway, and each step has specific and unique functions. A previous study has shown that this N-terminal domain name (NTD) of the v-SNARE docking to the t-SNARE is the rate-limiting step, which suggests that VAMP2 plays a crucial role in membrane fusion, mediating protein trafficking and the secretion of physiological mediators [18,19]. The role of VAMP2 is usually of vital importance in other processes in addition to membrane Docetaxel (Taxotere) fusion and vesicular transport. In a previous study, Yuki is still unknown. In the present study, we described the cloning, expression, and characterization of SjVAMP2, including the distribution of the protein in that were managed and released from Oncomelania hupensis snails infected with a Chinese mainland strain of restriction site underlined) as the sense primer and 5′-GTTCTCGAGTCACTGAGTAGCACTTCCA-3′ (restriction site underlined) as the antisense primer. PCR amplification was conducted under the following conditions: 1 min hold at 94C, 30 cycles of 94C for 30 s, 56.5C for 1 min, and 72C for 1min, followed by a hold for 10 min at 72C to ensure full.
