The presence of β-lactamases (e.g., PDC-3) that have naturally evolved and acquired the ability to break down β-lactam antibiotics ceftazidime ceftolozane) leads highly resistant potentially lethal Pseudomonas aeruginosa infections. We show wild-type PDC-3 β-lactamase forms an acyl enzyme complex with ceftazidime, but it cannot accommodate structurally similar ceftolozane has a longer R2 side chain increased basicity. A single amino acid substitution from glutamate lysine at position 221 in...

Understanding allostery in enzymes and tools to identify it offer promising alternative strategies inhibitor development. Through a combination of equilibrium nonequilibrium molecular dynamics simulations, we allosteric effects communication pathways two prototypical class A β-lactamases, TEM-1 KPC-2, which are important determinants antibiotic resistance. The simulations reveal operating over distances 30 Å or more. Propagation the signal occurs through cooperative coupling loop dynamics....

The D614G mutation in the Spike protein of SARS-CoV-2 has effectively replaced early pandemic-causing variant. Using pseudotyped lentivectors, we confirmed that aspartate replacement by glycine position 614 is markedly more infectious. Molecular modelling suggests G614 facilitates transition towards an open state protein. To explain epidemiological success D614G, analysed evolution 27,086 high-quality genome sequences from GISAID. We observed striking coevolution with P323L viral polymerase....

The mitogen-activated protein kinase (MAPK) p38α is a central component of signaling in inflammation and the immune response is, therefore, an important drug target. Little known about molecular mechanism its activation by double phosphorylation from MAPK kinases (MAP2Ks), because challenge trapping transient dynamic heterokinase complex. We applied multidisciplinary approach to generate structural model complex with MAP2K, MKK6, understand mechanism. Integrating cryo-electron microscopy...

Abstract The Epidermal Growth Factor Receptor (EGFR) is frequently found to be mutated in non-small cell lung cancer. Oncogenic EGFR has been successfully targeted by tyrosine kinase inhibitors, but acquired drug resistance eventually overcomes the efficacy of these treatments. Attempts surmount this therapeutic challenge are hindered a poor understanding how and why cancer mutations specifically amplify ligand-independent auto-phosphorylation signals enhance survival amplification related...

Kotzabasaki <italic>et al.</italic> use a multiscale computational approach to investigate the microscopic behaviour of gemcitabine stored in (OH)-IRMOF-74-III. The principles can be exploited for nano-carrier screening purposes prior experimental investigation.

Phosphoinositide 3-kinase alpha (PI3Kα) is involved in fundamental cellular processes including cell proliferation and differentiation frequently mutated human malignancies. One of the most common mutations E545K, which results an amino acid substitution opposite charge. It has been recently proposed that this oncogenic charge-reversal mutation, interactions between protein catalytic regulatory subunits are abrogated, resulting loss regulation constitutive PI3Kα activity, can lead to...

The rise of multi-drug resistance in bacterial pathogens is one the grand challenges facing medical science. A major concern speed development β-lactamase-mediated Gram-negative species, thus putting at risk efficacy most recently approved antibiotics and inhibitors, including carbapenems avibactam, respectively. New strategies to overcome are urgently required, which will ultimately be facilitated by a deeper understanding mechanisms that regulate function β-lactamases such as Klebsiella...

Mutations within the kinase domain of epidermal growth factor receptor (EGFR) are common oncogenic driver events in non-small cell lung cancer. Although activation EGFR normal cells is primarily driven by growth-factor-binding-induced dimerization, mutations on different exons have been found to affect equilibrium between its active and inactive conformations giving rise growth-factor-independent activation. Using molecular dynamics simulations combined with enhanced sampling techniques, we...

Non-structural protein 1 (Nsp1) is a main pathogenicity factor of α- and β-coronaviruses. Nsp1 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) suppresses the host gene expression by sterically blocking 40S ribosomal subunits promoting mRNA degradation. This mechanism leads to downregulation translation-mediated innate immune response in cells, ultimately mediating observed evasion capabilities SARS-CoV-2. Here, combining extensive molecular dynamics simulations, fragment...

The mechanism by which the cancer-causing E545K mutation may lead to PI3Kα activation is described in atomic-level detail.

Galectins are soluble β-D-galactoside-binding proteins whose implication in cancer progression and disease outcome makes them prominent targets for therapeutic intervention. In this frame, the development of small inhibitors that block selectively activity galectins represents an important strategy therapy which is, however, still relatively underdeveloped. To end, we designed here a rationally efficiently novel diglycosylated compound, characterized by selenoglycoside bond presence...

The sperm-specific channel CatSper (cation of sperm) controls the intracellular Ca2+ concentration ([Ca2+]i) and plays an essential role in sperm function. It is mainly activated by steroid progesterone (P4) but also promiscuously a wide range synthetic physiological compounds. These compounds include diverse steroids whose action on so far still controversial. To investigate effect these function, we developed high-throughput screening (HTS) assay to measure changes [Ca2+]i human screened...

Abstract The D614G mutation of the Spike protein is thought to be relevant for SARS-CoV-2 infection. Here we report biological and epidemiological aspects this mutation. Using pseudotyped lentivectors, were able confirm that G614 variant markedly more infectious than ancestral D614 variant. We demonstrate by molecular modelling replacement aspartate glycine in position 614 facilitates transition towards an open state protein. To understand whether increased infectivity explains its success,...

Phosphorylation is a fundamental mechanism in eukaryotic cells that allows signals to propagate. Kinases orchestrate this process by phosphorylating proteins modulate their activity. Our work reveals the architecture of complex between two key players mitogen-activated protein kinase signaling pathway, p38α and MKK6, while also giving an idea what happens when come together.

A cancer-associated missense mutation in the nuclear receptor RXRα acts by allosteric mechanisms and impacts differently activity of its dimers, depending on dimerization partner.

Abstract Non-structural protein 1 (Nsp1) is a main pathogenicity factor of α - and β -coronaviruses. Nsp1 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) suppresses the host gene expression by sterically blocking 40S ribosomal subunits promoting mRNA degradation. This mechanism leads to downregulation translation-mediated innate immune response in cells, ultimately mediating observed evasion capabilities SARS-CoV-2. Here, combining extensive Molecular Dynamics simulations,...

Abstract The MAP kinase p38α is a central component of signalling in inflammation and the immune response is, therefore, an important drug target. Little known about molecular mechanism its activation by double-phosphorylation from MAP2Ks, due to challenge trapping transient dynamic hetero-kinase complex. Here, we applied multidisciplinary approach generate first structure complex with MAP2K MKK6 understand mechanism. Integrating cryo-EM MD simulations, HDX-MS cellulo experiments,...

Abstract The sperm-specific channel CatSper ( cat ion of sper m) controls the intracellular Ca 2+ concentration ([Ca ] i ) and plays an essential role in sperm function. It is mainly activated by steroid progesterone (P4) but also promiscuously a wide range synthetic physiological compounds. These compounds include diverse steroids whose action on so far still controversial. To investigate effect these function, we developed high-throughput-screening (HTS) assay to measure changes [Ca human...

Summary Epidermal growth factor receptor (EGFR) is central to cell in physiology and pathophysiologies, including non-small lung cancer (NSCLC). EGFR has been successfully targeted with tyrosine kinase inhibitor generations, but the missense secondary T766M mutation a common cause of resistance. Overcoming this therapeutic challenge hindered by poor understanding how dysregulates function leading tumor progression. Here we show that amplifies vivo exploiting newly discovered oligomer...

Abstract Epidermal growth factor receptor (EGFR) is frequently mutated in non-small cell lung cancer (NSCLC) where it has been successfully targeted with tyrosine kinase inhibitors, but these are eventually overcome by drug resistance. Overcoming this therapeutic challenge hindered the poor understanding of how wild type (WT)-EGFR elicits ligand-independent signals essential for homeostasis, which then hijacked mutations. Ligand-independent signaling cannot be explained autoinhibited...