Vasoactive intestinal peptide (VIP) is implicated in many physiological and pathophysiological processes, and its receptors are promising targets for the development of new drugs. The human VPAC1 receptor for VIP and pituitary adenylate cyclase-activating polypeptide is a class II G protein coupled receptor. The N-terminal ectodomain (N-ted) of the VPAC1 receptor is a major VIP binding site. To determinate the high resolution structure of the VPAC1 receptor N-ted, large quantities of purified recombinant N-ted produced are required. The N-ted sequence (31-144), which is fused to thioredoxin protein and 6xHis tag, was expressed into Origami Escherichia coli strain. Purification of recombinant N-ted using Ni-NTA affinity column associated to Nu-polyacrylamide gel electrophoresis analysis reveals the presence of one single band of Mw 19,000 corresponding to the purified recombinant N-ted. The purified N-ted was able to recognize VIP and the selective antagonist PG96-269. About 5-10 mg of functional purified protein/liter of bacterial culture is currently produced. This is a crucial step to determine the structure of functional human VPAC1 receptor N-ted by nuclear magnetic resonance.
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Vasoactive intestinal peptide (VIP) is a neuropeptide involved in tumor growth and immune modulating functions. Previous research indicated that a VIP antagonist (VIPhyb) enhances T-cell activation and induces T-cell-dependent anti-leukemic activity in mice. We created a combinatorial library of VIPhyb C-terminal sequence variations to develop a more potent VIP-receptor (VIP-R) antagonist, hypothesizing that specific amino acid substitutions would improve receptor binding and plasma stability.
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January 2024
Departments of Animal Sciences, North Dakota State University, Fargo, ND 58108-6050, USA.
Article Synopsis
- - The study investigated how administering vasoactive intestinal polypeptide (VIP) affects inflammation and the expression of intestinal tight junction mRNA in lambs on grain-based diets, comparing two groups: one given VIP and another given saline (control).
- - Although there were no significant differences in tight junction mRNA expressions across intestinal regions, VIP-treated lambs showed increased levels of anti-inflammatory cytokines and lower lipopolysaccharide (LPS) concentrations, indicating reduced inflammation.
- - Overall, while VIP did not appear to change tight junction mRNA levels, the treatment seemed to lower inflammation by decreasing LPS levels in lambs fed grain diets.
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