TAT-tagging of VIP exerts positive allosteric modulation of the PAC1 receptor and enhances VIP neuroprotective effect in the MPTP mouse model of Parkinson's disease.

Background: The cationic Arginine-rich peptide (CARP) TAT had been tagged at the C-terminal end of the vasoactive intestinal peptide (VIP) to construct VIP-TAT in order to improve traversing ability. Interestingly, it was found that TAT may bind the positive allosteric modulation (PAM) site of the N-terminal extracellular domain of neuropeptide receptor PAC1 (PAC1-EC1), imitating the C-terminus part of pituitary adenylate cyclase-activating polypeptide (PACAP) PACAP(28-38) fragment.
Methods: To test this hypothesis, we addressed the neuroprotective effects of VIP, VIP-TAT and PACAP38 in Parkinson's Disease (PD) cellular and mouse models. We also analyzed the peptides affinity for PAC1 and their ability to activate it.
Results: VIP-TAT had in vitro and in vivo neuroprotective effects much efficient than VIP in PD cellular and mouse models. The isothermal titration calorimetry (ITC) and competition binding bioassays confirmed that TAT binds PAC1-EC1 at the same site as PACAP(28-38). The cAMP experiments showed TAT-VIP results in a higher activation potency of PAC1 than VIP alone.
Conclusions: The correlation of the peptides cationic properties with their affinity for PAC1 and their ability to activate the receptor, indicated that electrostatic interactions mediate the binding of TAT to the PAM domain of the PAC1-EC1, which induces the conformational changes of PAC1-EC1 required to promote the subsequent structural interaction and activation of the receptor with VIP.
General Significance: VIP-TAT has some potency for the development of a novel drug targeting neurodegenerative diseases.
Competing Interests: Declaration of Competing Interest All authors have read and approved this version of the manuscript, due care has been taken to ensure the integrity of the work. No original data has been published or submitted elsewhere. No conflict interest exists in the submission of this manuscript.
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