Ricerc@Sapienza

The notion that protein function is allosterically regulated by structural or dynamic changes in proteins has been extensively investigated in several protein domains in isolation. In particular, PDZ domains have represented a paradigm for these studies, despite providing conflicting results. Furthermore, it is still unknown how the association between protein domains in supramodules, consitituting so-called supertertiary structures, affects allosteric networks.

Although more than 75% of the proteome is composed of multidomain proteins, current knowledge of protein folding is based primarily on studies of isolated domains. In this work, we describe the folding mechanism of a multidomain tandem construct comprising two distinct covalently bound PDZ domains belonging to a protein called Whirlin, a scaffolding protein of the hearing apparatus. In particular, via a synergy between NMR and kinetic experiments, we demonstrate the presence of a misfolded intermediate that competes with productive folding.

The Envelope protein (E) is one of the four structural proteins encoded by the genome of SARS-CoV and SARS-CoV-2 Coronaviruses. It is an integral membrane protein, highly expressed in the host cell, which is known to have an important role in Coronaviruses maturation, assembly and virulence. The E protein presents a PDZ-binding motif at its C-terminus. One of the key interactors of the E protein in the intracellular environment is the PDZ containing protein PALS1.

SH2 domains are common protein interaction domains able to recognize short aminoacidic sequences presenting a phosphorylated tyrosine (pY). In spite of their fundamental importance for cell physiology there is a lack of information about the mechanism by which these domains recognize and bind their natural ligands. The N-terminal SH2 (N-SH2) domain of PI3K mediates the interaction with different scaffolding proteins and is known to recognize a specific pY-X-X-M consensus sequence.

SH2 domains are protein domains that modulate protein–protein interactions through a specific interaction with sequences containing phosphorylated tyrosines. In this work, we analyze the folding pathway of the C‐terminal SH2 domain of the p85 regulatory subunit of the protein PI3K, which presents a proline residue in a cis configuration in the loop between the βE and βF strands. By employing single and double jump folding and unfolding experiments, we demonstrate the presence of an on‐pathway intermediate that transiently accumulates during (un)folding.

Noonan syndrome (NS) is a genetic disorder caused by the hyperactivation of the RAS-MAPK molecular pathway. About 50% of NS cases are caused by mutations affecting the SHP2 protein, a multi-domain phosphatase with a fundamental role in the regulation of the RAS-MAPK pathway. Most NS-causing mutations influence the stability of the inactive form of SHP2. However, one NS-causing mutation, namely T42A, occurs in the binding pocket of the N-SH2 domain of the protein.

Although the vast majority of the human proteome is represented by multi-domain proteins, the study of multi-domain folding and misfolding is a relatively poorly explored field. The protein whirlin is a multi-domain scaffolding protein expressed in the inner ear. It is characterized by the presence of tandem repeats of PDZ domains. The first two PDZ domains of whirlin (PDZ1 and PDZ2 - namely P1P2) are structurally close and separated by a disordered short linker. We recently described the folding mechanism of the P1P2 tandem.

The paper investigated the effect of a physical surface abrasion of goji berries on drying kinetics and the evolution of phenolic compounds at 323, 333, and 343 K. A diffusion model was developed to describe the drying process. The effective diffusion coefficient estimated by the model ranged from 7.5 × 10–9 to 4.2 × 10–8 m2/s for cylinder and 2.0 × 10–9 to 1.15 × 10–8 m2/s for sphere for untreated samples (UTR). Higher values were found for treated samples (TR) (from 2.50 × 10–8 to 1.20 × 10–7 m2/s for cylinder and 8.0 × 10–9 to 2.70 × 10–8 m2/s for sphere).

Chitosan is very effective in removing metal ions through their adsorption. A preliminary investigation of the adsorption of chromium(III) by chitosan was carried out by means of batch tests as a function of contact time, pH, ion competition, and initial chromium(III) concentration. The rate of adsorption was rather rapid (t1/2 < 18 min) and influenced by the presence of other metal ions. The obtained data were tested using the Langmuir and Freundlich isotherm models and, based on R2 values, the former appeared better applicable than the latter.

The reactivity between NO and the oxidized form of a short-distance dinuclear Cu-ZSM-5 catalyst (ZCu2O) was investigated. ZCu2O, which contains the [Cu–O–Cu]2+bridge coordinated at the opposite T11 positions of the M6 ring of ZSM-5, is obtained by the spin-forbidden decomposition of N2O on the reduced form of the catalyst, ZCu2, with an activation energy of about 18 kcal mol−1. The further addition of NO to the [Cu–O–Cu]2+unit of ZCu2O occurs in the doublet state without activation energy and gives NO2.