title: An overview of the sequence features of N- and C-terminal segments of the human chemokine receptors.
abstract: Chemokine receptors play a crucial role in the cellular signaling enrolling extracellular ligands chemotactic proteins which recruit immune cells. They possess seven trans-membrane helices, an extracellular N-terminal region with three extracellular hydrophilic loops being important for search and recognition of specific ligand(s), and an intracellular C-terminal region with three intracellular loops that couple G-proteins. Although the functional aspects of the terminal segments of the extra-and intra-cellular G proteins are universally identified, the molecular basis on which they rest are still unclear because they are not definable by means of X-rays due to their high mobility and are not easy to study in the membrane. The purpose of this work is to define which physical-chemical properties of the terminal segments of the human chemokine receptors are at the basis of their functional mechanisms. Therefore, we have evaluated their physical-chemical properties in terms of amino acid composition, local flexibility, disorder propensity, net charge distribution and putative sites of post-translational modifications. Our results support the conclusion that all 19 C-terminal and N-terminal segments of human chemokine receptors are very flexible due to the systematic presence of intrinsic disorder. Although, the purpose of this plasticity clearly appears that of controlling and modulating the binding of ligands, we provide evidence that the overlap of linearly charged stretches, intrinsic disorder and post-translational modification sites, consistently found in these motives, is a necessary feature to exert the function. The role of the intrinsic disorder has been discussed considering the structural information coming from intrinsically disordered model compounds which support the view that the chemokine terminals have to be considered as strong polyampholytes or polyelectrolytes where conformational ensembles and structural transitions between them are modulated by charge fraction variations. Also the role of post-translational modifications has been found coherent with this view because, changing the charge fraction, they guide structural transitions between ensembles. Moreover, we have also considered our results from an evolutionary point of view in order to understand if the features found in humans were also present in other species. Our data evidenced that the structural features of the human terminals of the chemokine receptors were shared and evolutionarily conserved particularly among mammals. This means that the various organisms not only tolerate but select intrinsic disorder for the terminal regions of their receptors, reflecting constraints that point to molecular recognition. In conclusion the terminal segments of chemokine receptors must be considered as strong polyampholytes where the charge fraction variations induced by post-translational modifications are the driving physico-chemical feature able to adapt the conformations of the terminal segments to their functions.
authors.affiliation: Doctorate in Computational Biology, Second University of Naples, Naples, Italy; Department of Biochemistry, Biophysics and General Pathology, Second University of Naples, Naples, Italy.
authors.forename: Raffaele
authors.initials: R
authors.lastname: Raucci
authors.affiliation: Centro Ricerche Oncologiche di Mercogliano, Istituto Nazionale per lo studio e la cura dei tumori "Fondazione G. Pascale" - IRCCS, Italy. Electronic address: s.costantini@istitutotumori.na.it.
authors.forename: Susan
authors.initials: S
authors.lastname: Costantini
authors.affiliation: Centro Ricerche Oncologiche di Mercogliano, Istituto Nazionale per lo studio e la cura dei tumori "Fondazione G. Pascale" - IRCCS, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Castello
authors.affiliation: Doctorate in Computational Biology, Second University of Naples, Naples, Italy; Department of Biochemistry, Biophysics and General Pathology, Second University of Naples, Naples, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Colonna
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D064267:Intrinsically Disordered Proteins / Q000737:chemistry; D010802:Phylogeny; D011494:Protein Kinases / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D019707:Receptors, Chemokine / Q000737:chemistry*; D020539:Sequence Analysis, Protein
chemical_list: D064267:Intrinsically Disordered Proteins; D019707:Receptors, Chemokine; D011494:Protein Kinases
keywords: Chemokine receptors evolution; Conformational ensemble; Human chemokine receptors; Intrinsic protein disorder; Polyelectrolytes