title: Emergence of structure through protein-protein interactions and pH changes in dually predicted coiled-coil and disordered regions of centrosomal proteins.
abstract: Human centrosomal proteins show a significant, 3.5 fold, bias to be both unstructured and coiled-coils with respect to generic human proteins, based on results from state of the art bioinformatics tools. We hypothesize that this bias means that these proteins adopt an ensemble of disordered and partially helical conformations, with the latter becoming stabilized when these proteins form complexes. Characterization of the structural properties of 13 peptides from 10 different centrosomal proteins ranging in size from 20 to 61 residues by biophysical methods led us to confirm our hypothesis in most cases. Interestingly, the secondary structure adopted by most of these peptides becomes stabilized at acidic pH and it is concentration dependent. For two of them, PIK3R1(453-513) and BRCA1(1253-1273), we observed not only the stabilization of helical structure through self-association, but also the presence of β-structures linked to the formation of high molecular weight oligomers. These oligomers are the predominant forms detected by CD, but unobservable by liquid state NMR. BRCA1(1397-1424) and MAP3K11(396-441) populate helical structures that can also self-associate at pH3 through oligomeric species. Four peptides, derived from three proteins, namely CCNA2(103-123), BRCA1(1253-1273), BRCA1(1397-1424) and PIK3R1(453-513), can form intermolecular associations that are concomitant with alpha or beta structure stabilization. The self-phosphorylation previously described for the kinase NEK2 did not lead to any stabilization in the peptide's structure of NEK2(303-333), NEK2(341-361), and NEK2(410-430). Based on these results, obtained from a series of peptides derived from a significant number of different centrosomal proteins, we propose that conformational polymorphism, modulated by intermolecular interactions is a general property of centrosomal proteins.
authors.affiliation: Instituto de Química Física "Rocasolano", CSIC, Serrano 119, 28006 Madrid, Spain. Electronic address: mtrevino@iqfr.csic.es.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Treviño
authors.affiliation: Instituto de Química Física "Rocasolano", CSIC, Serrano 119, 28006 Madrid, Spain. Electronic address: mgarcia@iqfr.csic.es.
authors.forename: M Flor
authors.initials: MF
authors.lastname: García-Mayoral
authors.affiliation: Instituto de Química Física "Rocasolano", CSIC, Serrano 119, 28006 Madrid, Spain. Electronic address: majimenez@iqfr.csic.es.
authors.forename: M Ángeles
authors.initials: MÁ
authors.lastname: Jiménez
authors.affiliation: Centro de Biología Molecular Severo Ochoa (CBMSO), CSIC-UAM, Cantoblanco, Madrid, Spain. Electronic address: ubastolla@cbm.csic.es.
authors.forename: Ugo
authors.initials: U
authors.lastname: Bastolla
authors.affiliation: Instituto de Química Física "Rocasolano", CSIC, Serrano 119, 28006 Madrid, Spain. Electronic address: mbruix@iqfr.csic.es.
authors.forename: Marta
authors.initials: M
authors.lastname: Bruix
keywords: CD; Centrosome; Coiled-coil; Disordered regions; NMR; Peptides