Physiology and Pharmacology “Vittorio Erspamer”, Sapienza University of Rome, Piazzale Aldo Moro five, 00185 Rome, Italy; [email protected] (D.M.); [email protected] (M.V.) Division of Biochemical Sciences “Alessandro Rossi Fanelli”, Sapienza University of Rome, Piazzale Aldo Moro five, 00185 Rome, Italy; [email protected] Correspondence: [email protected] (R.L.); [email protected] (R.M.)Citation: Lattanzi, R.; Maftei, D.; Vincenzi, M.; Fullone, M.R.; Miele, R. Identification and Characterization of a new Splicing Variant of Prokineticin 2. Life 2022, 12, 248. doi.org/10.3390/ life12020248 Academic Editor: Kristian Stromgaard Received: 23 December 2021 Accepted: 31 January 2022 Published: 7 February 2022 Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.Abstract: Prokineticin 2 (PROK2) is usually a secreted bioactive peptide that regulates many different biological responses through two GPCRs, the prokineticin receptors (PROKRs). The aim of this study was to characterize a brand new alternatively spliced solution from the prok2 gene consisting of 4 exons. The 40-amino acid peptide, designated PROK2C, is encoded by exon 1 and exon 4, and its expression was detected in the hippocampus and spinal cord of mice.HMGB1/HMG-1 Protein web PROK2C was expressed within a heterologous system, Pichia pastoris, and its binding specificity to the amino-terminal regions of PROKR1 and PROKR2 was investigated by GST pull-down experiments. Moreover, the introduction on the unnatural amino acid p-benzoyl-L-phenylalanine working with amber codon suppression technologies demonstrated the part of tryptophan at position 212 of PROKR2 for PROK2C binding by photoactivatable cross-linking. The functional significance of this new isoform was determined in vivo by nociceptive experiments, which showed that PROK2C elicits strong sensitization of peripheral nociceptors to painful stimuli. To be able to analyze the induction of PROK2C signal transduction, STAT3 and ERK phosphorylation levels were determined in mammalian CHO cells expressing PROKR1 and PROKR2. Our data show by in vivo and in vitro experiments that PROK2C can bind and activate each prokineticin receptors. Search phrases: option splicing; prokineticin two; prokineticin 2 splice variant; prokineticin receptors1.Adiponectin/Acrp30 Protein Biological Activity Introduction Alternative splicing (AS) is definitely an crucial mechanism in gene modulation.PMID:23812309 It makes it possible for a single gene to create quite a few unique mRNAs with different exon compositions and lengths that can code for many forms from the corresponding proteins. Most eukaryotic protein-coding genes involved in developmental processes and regulation of cell proliferation have numerous transcriptional isoforms [1,2]. AS can affect mRNA localization, stability, and translation or change the reading frame, resulting in distinctive protein isoforms with diverse functions, localizations, or both [3,4]. AS is very regulated by various regulatory components of splicing that modulate splice web-site selection and spliceosome assembly. Any alteration in the splicing mechanism can impact the maturation of mRNA and consequently the formation of functional proteins, causing many pathological states such as neurodegenerative ailments and inflammatory, immune, and metabolic disorders [5,6]. Prokineticin two (PROK2) is a chemokine-like protein found in all evolutionary scales, such as reptiles, amphibians, mammals, and humans, and is characterized by finely regu.