The protease encoded by the TMPRSS2 gene facilitates viral infections and has been implicated in pathogenesis of SARS-CoV-2. We analyzed sequence correlated protein variants with clinical features a cohort 1177 patients affected COVID-19 Italy. Nine relatively common (allele frequency > 0.01) six missense which may affect activity according to PolyPhen-2 HumVar-trained mode were identified. Among them, p.V197M (p.Val197Met) (rs12329760) emerges as variant that deleterious effect on...

Thermal shift assay can be used for the high‐throughput screening of pharmacological chaperones. These drugs are small molecules that bind a mutant protein and stabilize it. We demonstrated robustness, reproducibility versatility method using two in clinical trial Fabry or Pompe disease, Deoxygalactonojirimycin N‐Butyldeoxynojirimycin, their target enzymes, lysosomal alpha‐galactosidaseA alpha‐glucosidase, as test cases. assessed influence solvents scanning rate on measures. showed value is...

Fabry disease is a lysosomal storage caused by mutations in the GLA gene that encodes alpha-galactosidase (AGAL). The causes abnormal globotriaosylceramide (Gb3) lysosomes. Variants responsible for genotypic spectrum of include abolish enzymatic activity and those cause protein instability. latter can be successfully treated with small molecules either bind stabilize AGAL or indirectly improve its cellular activity. This paper describes first attempt to reposition curcumin, nutraceutical,...

Fabry disease is caused by a deficiency of lysosomal alpha galactosidase and has very large genotypic phenotypic spectrum. Some patients who carry hypomorphic mutations can benefit from oral therapy with pharmacological chaperone. The drug requires precise regimen because it reversible inhibitor alpha-galactosidase. We looked for molecules that potentiate this chaperone, among drugs have already been approved other diseases. tested candidate in fibroblasts derived patient carrying deletion...

Enzyme replacement therapy is the only therapeutic option for Fabry patients with completely absent AGAL activity. However, treatment has side effects, costly, and requires conspicuous amounts of recombinant human protein (rh-AGAL). Thus, its optimization would benefit welfare/health services (i.e., society at large). In this brief report, we describe preliminary results paving way two possible approaches: i. combination enzyme pharmacological chaperones; ii. identification interactors as...

Many bioactive peptides, presenting an unstructured conformation in aqueous solution, are made resistant to degradation by posttranslational modifications. Here, we describe how molecular oligomerization solution can generate a still unknown transport form for amphipathic which is more compact and proteases than forms related any possible monomer. This phenomenon emerged from 3D structure, function, properties of distinctin, heterodimeric antimicrobial compound consisting two peptide chains...

Type I disorders of glycosylation (CDG), the most frequent which is phosphomannomutase 2 (PMM2-CDG), are a group diseases causing incomplete N-glycosylation proteins. PMM2-CDG an autosomal recessive disease with large phenotypic spectrum, and associated mutations in PMM2 gene. The biochemical analysis mutants does not allow precise genotype–phenotype correlation for PMM2-CDG. very tolerant to missense loss function mutations, suggesting that partial deficiency activity might be beneficial...

The most common glycosylation disorder is caused by mutations in the gene encoding phosphomannomutase2, producing a disease still without cure. Phosphomannomutase2, homodimer which each chain composed of two domains, requires bisphosphate sugar (either mannose or glucose) as activator, opening possible drug design path for therapeutic purposes. crystal structure human however, lacks bound substrate and key active site loop. To speed up discovery, we present here first structural model to...

A large number of mutations causing PMM2-CDG, which is the most frequent disorder glycosylation, destabilize phosphomannomutase2. We looked for a pharmacological chaperone to cure starting from structure natural ligand phosphomannomutase2, α-glucose-1,6-bisphosphate. The compound, β-glucose-1,6-bisphosphate, was synthesized and characterized via 31P-NMR. β-glucose-1,6-bisphosphate binds its target enzyme in silico. binding induces conformational change that predicted by program PELE...

The 32 amino acid hormone human calcitonin was studied at pH 3.7 and 7.4 by multidimensional NMR spectroscopy in sodium dodecyl sulfate micelles 310K. secondary structure obtained from nuclear Overhauser enhancement (NOESY), 3JHNα coupling constants, slowly exchanging amide data. Three-dimensional structures consistent with data were generated using distance geometry calculations. A set of 265 interproton distances derived NOESY experiments, hydrogen-bond constraints exchange, constants...

Salmon calcitonin (sCT) forms an amphipathic helix in the region 9-19, with C-terminal decapeptide interacting (Amodeo, P., Motta, A., Strazzullo, G., Castiglione Morelli, M. A. (1999) J. Biomol. NMR 13, 161-174). To uncover structural requirements for hormone bioactivity, we investigated several sCT analogs. They were designed so as to alter length of central by removal and/or replacement flanking residues and selectively mutating or deleting inside helix. The content was assessed circular...

Fabry disease is a rare disorder caused by large variety of mutations in the gene encoding lysosomal alpha-galactosidase. Many these are unique to individual families. can be treated with enzyme replacement therapy, but promising novel strategy relies on small molecules, so called "pharmacological chaperones", which administered orally. Unfortunately only 42% genotypes respond pharmacological chaperones. A procedure predict responsive chaperones has been recently proposed. The method uses...

The most frequent disorder of glycosylation is due to mutations in the gene encoding phosphomannomutase2 (PMM2-CDG). For this disease, which autosomal and recessive, there no cure at present. Most patients are composite heterozygous carry one allele an inactive mutant, R141H, a hypomorphic mutant. Phosphomannomutase2 dimer. We reproduced heterozygosity vitro by mixing R141H either with wild type protein or common mutant F119L compared quaternary structure, activity stability heterodimeric...

Personalized therapies are required for Fabry disease due to its large phenotypic spectrum and numerous different genotypes. In principle, missense mutations that do not affect the active site could be rescued with pharmacological chaperones. At present chaperones bind couple a stabilizing effect, which is required, an inhibitory deleterious. By in silico docking we identified allosteric hot-spot ligand binding where drug-like compound, 2,6-dithiopurine, binds preferentially....

The most common cause of human congenital disorders glycosylation (CDG) are mutations in the phosphomannomutase gene PMM2, which affect protein N -linked glycosylation. yeast SEC53 encodes a homolog PMM2 . We evolved 384 populations harboring one two human-disease-associated alleles, sec53- V238M and sec53 -F126L, or wild-type find that after 1000 generations, compensate for slow-growth phenotype associated with alleles. Through whole-genome sequencing we identify compensatory mutations,...

Abstract Background The pharmacological chaperones therapy is a promising approach to cure genetic diseases. It relies on substrate competitors used at sub-inhibitory concentration which can be administered orally, reach difficult tissues and have low cost. Clinical trials are currently carried out for Fabry disease, lysosomal storage disorder caused by inherited mutations of alpha-galactosidase. Regrettably, not all genotypes respond these drugs. Results We collected the experimental data...

Irreversible aggregation limits bioavailability and therapeutic activity of protein-based drugs. Here we show that an aggregation-resistant mutant can be engineered by structural homology with a non-amyloidogenic analogue the variant may act as inhibitor. This strategy has successfully been applied to amyloidogenic human calcitonin (hCT). Including only five residues from salmon (sCT), obtained variant, polar (phCT), whose solution structure was shown CD, NMR, calculations practically...