The human somatic angiotensin converting enzyme (ACE) contains two homologous domains, each bearing a zinc-dependent active site. All of the synthetic inhibitors this used in clinical applications interact with these sites to similar extent. Recently, several lines evidence have suggested that N-terminal site ACE might be involved specific hydrolysis some important physiological substrates, like Acetyl-Seryl-Aspartyl-Lysyl-Proline, negative regulator hematopoietic stem cell differentiation...

Abstract Recent data have pointed to TNALP as a therapeutic target for soft-tissue ossification abnormalities. Here, we used mutagenesis, kinetic analysis, and computer modeling identify the residues important binding of known ALP inhibitors active site. These will enable drug design efforts aimed at developing improved specific use. Introduction: We shown previously that genetic ablation tissue-nonspecific alkaline phosphatase (TNALP) function leads amelioration in mouse models...

Disulfide-rich proteins or DRPs are versatile bioactive compounds that encompass a wide variety of pharmacological, therapeutic, and/or biotechnological applications. Still, the production in sufficient quantities is major bottleneck for their complete structural functional characterization. Recombinant expression such small containing multiple disulfide bonds bacteria E. coli considered difficult and general methods protocols, particularly on high throughput scale, limited. Here we report...

Barrier-to-autointegration factor (BAF), encoded by the BANF1 gene, is an abundant and ubiquitously expressed metazoan protein that has multiple functions during cell cycle. Through its ability to cross-bridge two double-stranded DNA (dsDNA), it favours chromosome compaction, participates in post-mitotic nuclear envelope reassembly essential for repair of large ruptures. BAF forms a ternary complex with proteins lamin A/C emerin, interaction defective patients recessive accelerated aging...

Somatic angiotensin I converting enzyme (ACE) contains two functional active sites. Up to now, most of the studies aimed at characterizing selectivity inhibitors toward ACE sites relied on use mutants containing a single site. By developing new fluorogenic synthetic substrates ACE, we demonstrated that inhibitor can be assessed directly by using somatic ACE. This useful screening approach led us discover some bradykinin potentiating peptides turned out selective C-domain The peptide...

Angiotensin-converting enzyme 2 (ACE2), a recently identified human homologue of angiotensin-converting enzyme, is zinc metallocarboxypeptidase which may play unique role in cardiovascular and renal function. Here we report the discovery potent selective inhibitors ACE2, have been by evaluating series phosphinic di- tripeptides general formula: Z-Xaa(PO2-CH2)YaaOH Ac-Zaa-Xaa(PO2-CH2)YaaOH. The most inhibitor this tripeptide that displays Ki value 0.4 nM toward ACE2 3 orders magnitude less...

Abstract Specific proteins present at telomeres ensure chromosome end stability, in large part through unknown mechanisms. In this work, we address how the Saccharomyces cerevisiae ORC-related Rif2 protein protects telomere. We show that small N-terminal BAT motif ( B locks A ddition of T elomeres) previously known to limit telomere elongation and Tel1 activity is also sufficient block NHEJ 5’ resection. The inhibits ability Mre11-Rad50-Xrs2 complex (MRX) capture DNA ends. It acts a direct...

RXPA380 (Cbz-PheΨ[PO2CH]Pro-Trp-OH) was reported recently as the first highly selective inhibitor of C-domain somatic angiotensin-converting enzyme (ACE), able to differentiate two active sites ACE by a selectivity factor more than 3 orders magnitude. The contribution each residue toward this remarkable evaluated studying several analogues RXPA380. This analysis revealed that both pseudo-proline and tryptophan residues in P1' P2' positions play critical role for C-domain. is not due...

Detection of structural motif residues in protein structures allows identification or functional similarity between proteins. In the field engineering, is essential to select scaffolds on which a can be transferred design new with given function. We describe here RASMOT-3D PRO webserver (http://biodev.extra.cea.fr/rasmot3d/) that performs systematic search 3D for set exhibiting particular topology. Comparison based Cα and Cβ atoms two steps: inter-atomic distances RMSD. takes input PDB file...

Abstract The dynamic coupling between a polarizable protein force field and particle‐based implicit solvent model is described. field, TCPEp, developed recently to simulate systems, characterized by reduced number of sites, with substantial gain in efficiency for an equal chemical accuracy. Polarizable Pseudo‐Particle (PPP) represents the macroscopic polarization induced dipoles placed on mobile Lennard‐Jones pseudo‐particles. solvent‐induced are sensitive solute electric but not each other,...

Abstract A revised and improved version of our efficient polarizable force‐field/coarse grained solvent combined approach (Masella, Borgis, Cuniasse, J. Comput. Chem. 2008, 29, 1707) is described. The pseudo‐particle model represents the macroscopic polarization by induced dipoles placed on mobile pseudo‐particles. In this study, we propose a new formulation energy term handling nonelectrostatic interactions among This now able to reproduce energetic structural response liquid water due...

Matrix metalloproteinase (MMP)-13 is one of the mammalian collagenases that play key roles in tissue remodelling and repair progression diseases such as cancer, arthritis, atherosclerosis, aneurysm. For collagenase to cleave triple helical collagens, structure has be locally unwound before hydrolysis, but this process not well understood. We report crystal structures catalytically inactive full-length human MMP-13(E223A) complex with peptides 14–26 aa derived from cleaved prodomain during...

Abstract Bacteriophage capsids constitute icosahedral shells of exceptional stability that protect the viral genome. Many display on their surface decoration proteins whose structure and function remain largely unknown. The protein pb10 phage T5 binds at centre 120 hexamers formed by major capsid protein. Here we determined 3D investigated its capsid-binding properties using NMR, SAXS, cryoEM SPR. Pb10 consists an α-helical domain Ig-like exposed to solvent. It with a remarkably high...

Here, we report the molecular engineering of nanobodies that bind with picomolar affinity to both SARS-CoV-1 and SARS-CoV-2 receptor-binding domains (RBD) are highly neutralizing. We applied deep mutational VHH72, a nanobody initially specific for RBD little cross-reactivity antigen. first identified all individual VHH substitutions increase binding then screened focused combinatorial libraries isolate engineered improved properties. The corresponding VHH-Fc molecules show high affinities...

The application of an ensemble-averaging (EA) protocol to highlight conformational variability and determine the interconverting conformations in NMR structure cyclopeptides is described. Most NMR-based studies reported literature rely on protocols that basically assume existence a single structure. This sometimes referred as one NOE (or ROE)/one distance hypothesis. In contrast, EA used this work relies model explicitly takes into account averaging data tests significancy results which very...

Among natural metalloenzymes, the facial two-histidines one-carboxylate binding motif (FTM) is a widely represented first coordination sphere present in active site of variety metalloenzymes. A PDB search revealed total 1685 structures bearing such FTMs bound to metal. Sixty statistically representative were selected and used as template for identification structurally characterized proteins these three amino acids propitious environment transition This geometrical superposition search,...

A new model to study proteinic systems including a many-body polarization and hydrogen bond energy contribution is presented. This represents an extension of earlier water [M. Masella J.-P. Flament, J. Chem. Phys. 107 9105 (1997)]. As in this model, the developed reproduce quantum computations on small molecular aggregates, and, first paper, we focus our efforts developing accurate potential describe interactions among all nonbonded atoms occurring proteins, those six cations biological...

Abstract A multiscale coarse‐grained approach able to handle efficiently the solvation of microscopic solutes in extended chemical environment is described. That compute readily and very long‐range solute/solvent electrostatic interactions, up 1‐μm scale, by considering a reduced amount computational resources. All required parameters are assigned reproduce available data concerning single ions. Such strategy makes it possible with good accuracy properties simple ion pairs solution (in...

Barrier-to-autointegration factor (BAF) is an essential component of the nuclear lamina. Encoded by BANF1, this DNA binding protein contributes to regulation gene expression, cell cycle progression, and integrity. A rare recessive BAF variant, Ala12Thr, causes premature aging syndrome, Néstor–Guillermo progeria syndrome (NGPS). Here, we report first dominant pathogenic Gly16Arg, identified in a patient presenting with progressive neuromuscular weakness. Although disease variants carry nearby...

We have developed a structure-based approach to the design of protein ligands. This is based on transfer functional binding motif amino acids, often referred as "hot spot", host able reproduce topology these residues. The scaffolds were identified by systematic in silico search Protein Data Bank for proteins possessing group residues similar that adopted reference ligand known 3D structure. In contrast previously reported studies, this independent particular secondary structure supporting...

Somatic angiotensin‐converting enzyme (ACE) contains two homologous domains, each bearing a functional active site. Studies on the selectivity of these ACE domains towards either substrates or inhibitors have mostly relied use mutants isolated ACE. To determine directly properties domain, working with wild‐type enzyme, we developed an approach based combined N‐domain‐selective and C‐domain‐selective fluorogenic substrates. With this approach, marked differences in substrate were revealed...