A5023885000- Monoclonal and Polyclonal Antibodies Research
- HER2/EGFR in Cancer Research
- Radiopharmaceutical Chemistry and Applications
- DNA Repair Mechanisms
- DNA and Nucleic Acid Chemistry
- RNA and protein synthesis mechanisms
- CAR-T cell therapy research
- Enzyme Structure and Function
- CRISPR and Genetic Engineering
- Antimicrobial Resistance in Staphylococcus
- Genomics and Chromatin Dynamics
- Protein Kinase Regulation and GTPase Signaling
- Bacterial biofilms and quorum sensing
- Receptor Mechanisms and Signaling
- Nanofabrication and Lithography Techniques
- Glycosylation and Glycoproteins Research
- Ubiquitin and proteasome pathways
- Bacterial Genetics and Biotechnology
- Cell Adhesion Molecules Research
- Toxin Mechanisms and Immunotoxins
- Antimicrobial Peptides and Activities
- Molecular Biology Techniques and Applications
- Protein Structure and Dynamics
- Biochemical and Structural Characterization
- Plant Molecular Biology Research
CIC bioGUNE
2015-2024
Euskadiko Parke Teknologikoa
2019
Institut de Chimie
2010
Institut de génétique et de biologie moléculaire et cellulaire
2010
Genomics (United Kingdom)
2010
Ikerbasque
2010
Spanish National Cancer Research Centre
2010
Université de Strasbourg
2010
Gobierno de Navarra
2008-2009
Agrobiotechnology Institute
2005-2009
The capacity of Staphylococcus aureus to form biofilms on host tissues and implanted medical devices is one the major virulence traits underlying persistent chronic infections. matrix in which S. cells are encased a biofilm often consists polysaccharide intercellular adhesin (PIA) or poly-N-acetyl glucosamine (PNAG). However, surface proteins capable promoting development absence PIA/PNAG exopolysaccharide have been described. Here, we used two-dimensional nano-liquid chromatography mass...
Abstract In eukaryotes, DNA polymerase δ (Pol δ) bound to the proliferating cell nuclear antigen (PCNA) replicates lagging strand and cooperates with flap endonuclease 1 (FEN1) process Okazaki fragments for their ligation. We present high-resolution cryo-EM structure of human processive Pol δ–DNA–PCNA complex in absence presence FEN1. is anchored one three PCNA monomers through C-terminal domain catalytic subunit. The core sits on top an open configuration while regulatory subunits project...
The biofilm formation capacity of Staphylococcus aureus clinical isolates is considered an important virulence factor for the establishment chronic infections. Environmental conditions affect S. aureus, indicating existence positive and negative regulators process. majority screening procedures identifying genes involved in development have been focused on whose presence essential In this report, we used random transposon mutagenesis systematic disruption all two-component systems to...
Staphylococcus aureus can establish chronic infections on implanted medical devices due to its capacity form biofilms. Analysis of the factors that assemble cells into a biofilm has revealed occurrence strains produce either polysaccharide intercellular adhesin/poly-N-acetylglucosamine (PIA/PNAG) exopolysaccharide- or protein-dependent biofilm. Examination influence matrix nature capacities embedded bacteria remained elusive, because natural strain readily converts between polysaccharide-...
Sequence-specific endonucleases recognizing long target sequences are emerging as powerful tools for genome engineering. These could be used to correct deleterious mutations or inactivate viruses, in a new approach molecular medicine. However, such applications highly demanding terms of safety. Mutations the human RAG1 gene cause severe combined immunodeficiency (SCID). Using I-CreI dimeric LAGLIDADG meganuclease scaffold, we describe here engineering series cleaving gene, including obligate...
The costimulation of immune cells using first-generation anti-4-1BB monoclonal antibodies (mAbs) has demonstrated anti-tumor activity in human trials. Further clinical development, however, is restricted by significant off-tumor toxicities associated with FcγR interactions. Here, we have designed an Fc-free tumor-targeted 4-1BB-agonistic trimerbody, 1D8N/CEGa1, consisting three single-chain variable fragments and anti-EGFR single-domain positioned extended hexagonal conformation around the...
The intrinsically disordered protein p15PAF regulates DNA replication and repair by binding to the proliferating cell nuclear antigen (PCNA) sliding clamp. We present structure of human p15PAF–PCNA complex. Crystallography NMR show central PCNA-interacting motif (PIP-box) tightly bound front-face PCNA. In contrast other proteins, also contacts inside of, passes through, PCNA ring. termini emerge at opposite faces ring, but remain protected from 20S proteasomal degradation. Both free...
Abstract Sliding clamps encircle DNA and tether polymerases other factors to the genomic template. However, molecular mechanism of clamp sliding on is unknown. Using crystallography, NMR dynamics simulations, here we show that human PCNA recognizes through a double patch basic residues within ring channel, arranged in right-hand spiral matches pitch B-DNA. We propose slides by tracking backbone via ‘cogwheel’ based short-lived polar interactions, which keep orientation invariant relative...
The redirection of T cell activity using bispecific antibodies is one the most promising cancer immunotherapy approaches currently in development, but it limited by cytokine storm-related toxicities, as well pharmacokinetics and tumor-penetrating capabilities current antibody formats. Here, we have engineered ATTACK (Asymmetric Tandem Trimerbody for Activation Cancer Killing), a novel cell-recruiting which combines three EGFR-binding single-domain (VHH; clone EgA1) with single CD3-binding...
Biofilm formation in Staphylococcus aureus is usually associated with the production of poly-N-acetylglucosamine (PNAG) exopolysaccharide, synthesized by proteins encoded icaADBC operon. PNAG a linear β-(1-6)-linked N-acetylglucosaminoglycan that has to be partially deacetylated and consequently positively charged order bacterial cell surfaces. Here, we investigated whether attachment surfaces mediated ionic interactions negative charge wall teichoic acids (WTAs), which represent most...
ABSTRACT Biofilm formation in Staphylococcus aureus is subject to phase variation, and biofilm-negative derivatives emerge sporadically from a biofilm-positive bacterial population. To date, the only known mechanism for generating biofilm phenotypic variation staphylococci reversible insertion/excision of IS 256 biofilm-essential genes. In this study, we present evidence suggesting that absence σ B transcription factor dramatically increases rate switching phenotype clinical isolate S....
Homing endonucleases recognize long target DNA sequences generating an accurate double-strand break that promotes gene targeting through homologous recombination. We have modified the homodimeric I-CreI endonuclease protein engineering to a specific sequence within human RAG1 gene. Mutations in produce severe combined immunodeficiency (SCID), monogenic disease leading defective immune response individuals, leaving them vulnerable infectious diseases. The structures of two engineered...
Abstract Heterotrimeric G proteins are quintessential signalling switches activated by nucleotide exchange on Gα. Although activation is predominantly carried out G-protein-coupled receptors (GPCRs), non-receptor guanine-nucleotide factors (GEFs) have emerged as critical molecules and therapeutic targets. Here we characterize the molecular mechanism of G-protein a family GEFs containing Gα-binding -activating (GBA) motif. We combine NMR spectroscopy, computational modelling biochemistry to...
Abstract Here, we describe a new strategy that allows the rapid and efficient engineering of mono multispecific trivalent antibodies. By fusing single-domain antibodies from camelid heavy-chain-only immunoglobulins (V HHs ) to N-terminus human collagen XVIII trimerization domain (TIE produced monospecific trimerbodies were efficiently secreted as soluble functional proteins by mammalian cells. The purified V HH -TIE trimeric in solution exhibited excellent antigen binding capacity....
The INhibitor of Growth (ING) family tumor suppressors regulates the transcriptional state chromatin by recruiting remodeling complexes to sites with histone H3 trimethylated at lysine 4 (H3K4me3). This modification is recognized plant homeodomain (PHD) present C-terminus five ING proteins. ING5 facilitates acetylation HBO1 complex, and also H4 MOZ/MORF complex. We show that forms homodimers through its N-terminal domain, which folds independently into an elongated coiled-coil structure....
Abstract Purpose: The induction of 4-1BB signaling by agonistic antibodies can drive the activation and proliferation effector T cells thereby enhance a T-cell–mediated antitumor response. Systemic administration anti-4-1BB–agonistic IgGs, although effective preclinically, has not advanced in clinical development due to their severe hepatotoxicity. Experimental Design: Here, we generated humanized EGFR-specific 4-1BB-agonistic trimerbody, which replaces IgG Fc region with human collagen...
PCNA is an essential factor for DNA replication and repair. It forms a ring shaped structure of 86 kDa by the symmetric association three identical protomers. The encircles acts as docking platform other proteins, most them containing Interaction Protein sequence (PIP-box). We have used NMR to characterize interactions with several proteins fragments in solution. binding PIP-box peptide cell cycle inhibitor p21 consistent crystal complex. A shorter binds reduced affinity but retains...
Here, we describe a new class of multivalent and multispecific antibody-based reagents for therapy. The molecules, termed "trimerbodies," use modified version the N-terminal trimerization region human collagen XVIII noncollagenous 1 domain flanked by two flexible linkers as trimerizing scaffold. By fusing single-chain variable fragments (scFv) with same or different specificity to both N- C-terminus scaffold domain, produced monospecific bispecific hexavalent molecules that were efficiently...
p15PAF is an oncogenic intrinsically disordered protein that regulates DNA replication and lesion bypass by interacting with the human sliding clamp PCNA. In absence of DNA, traverses PCNA ring via extended PIP-box contacts surface. Here, we probed atomic-scale structure p15PAF-PCNA-DNA ternary complexes. Crystallography MD simulations show that, when occupies two subunits homotrimer, within channel binds unoccupied subunit. The PCNA-bound in presence invariant, solution NMR confirms does...
Abstract chTOG is a conserved microtubule polymerase that catalyses the addition of tubulin dimers to promote growth. interacts with TACC3, member transforming acidic coiled-coil (TACC) family. Here we analyse their association using Xenopus homologues, XTACC3 (TACC3) and XMAP215 (chTOG), dissecting mechanism by which interaction promotes elongation during spindle assembly. Using SAXS, show TACC domain (TD) an elongated structure mediates C terminus XMAP215. Our data suggest one TD two...
Abstract Heterotrimeric G proteins are usually activated by the guanine-nucleotide exchange factor (GEF) activity of GPCRs. However, some non-receptor also GEFs. GIV (a.k.a Girdin) was first protein for which GEF ascribed to a well-defined sequence that directly binds Gαi. expression promotes metastasis and disruption its binding Gαi blunts pro-metastatic behavior cancer cells. Although this suggests inhibition Gαi-GIV interaction is promising therapeutic strategy, protein-protein...
4-1BB (CD137) is an inducible costimulatory receptor that promotes expansion and survival of activated T cells; IgG-based 4-1BB-agonistic monoclonal antibodies exhibited potent antitumor activity in clinical trials. However, the development those restricted by major off-tumor toxicities associated with FcγR interactions. We have recently generated EGFR-targeted trimerbody demonstrated strong did not induce systemic inflammatory cytokine secretion hepatotoxicity first-generation agonists....
The four member family of “Cyclin and Cystathionine β-synthase (CBS) domain divalent metal cation transport mediators”, CNNMs, are the least-studied mammalian magnesium mediators. CNNM4 is abundant in brain intestinal tract, its abnormal activity causes Jalili Syndrome. Recent findings show that suppression mice promotes malignant progression polyps linked to infertility. association with phosphatases regenerating liver, PRLs, abrogates Mg2+-efflux capacity, thus resulting an increased...