Miquel Vila-Perello

Miquel Vila-Perello

Barcelona, Catalonia, Spain
1K followers 500+ connections

Join to view profile

About

Experienced executive, project leader and researcher with a demonstrated history of…

Activity

Join now to see all activity

Experience

Education

Publications

  • Chemical tagging and customizing of cellular chromatin states using ultrafast trans-splicing inteins

    Nature Chemistry

    Other authors
    See publication

  • Structure of the branched intermediate in protein splicing

    Proceedings of the National Academy of Sciences

    Inteins are autoprocessing domains that cut themselves out of host proteins in a traceless manner. This process, known as protein splicing, involves multiple chemical steps that must be coordinated to ensure fidelity in the process. The committed step in splicing involves attack of a conserved Asn side-chain amide on the adjacent backbone amide, leading to an intein-succinimide product and scission of that peptide bond. This cleavage reaction is stimulated by formation of a branched…

    Inteins are autoprocessing domains that cut themselves out of host proteins in a traceless manner. This process, known as protein splicing, involves multiple chemical steps that must be coordinated to ensure fidelity in the process. The committed step in splicing involves attack of a conserved Asn side-chain amide on the adjacent backbone amide, leading to an intein-succinimide product and scission of that peptide bond. This cleavage reaction is stimulated by formation of a branched intermediate in the splicing process. The mechanism by which the Asn side-chain becomes activated as a nucleophile is not understood. Here we solve the crystal structure of an intein trapped in the branched intermediate step in protein splicing. Guided by this structure, we use protein-engineering approaches to show that intein-succinimide formation is critically dependent on a backbone-to-side-chain hydrogen-bond. We propose that this interaction serves to both position the side-chain amide for attack and to activate its nitrogen as a nucleophile. Collectively, these data provide an unprecedented view of an intein poised to carry out the rate-limiting step in protein splicing, shedding light on how a nominally nonnucleophilic group, a primary amide, can become activated in a protein active site.

    Other authors
    See publication

  • Induction of innate and adaptive immunity by delivery of poly dA:dT to dendritic cells

    Nature Chemical Biology

    Targeted delivery of antigens to dendritic cells (DCs) is a promising vaccination strategy. However, to ensure immunity, the approach depends on coadministration of an adjuvant. Here we ask whether targeting of both adjuvant and antigen to DCs is sufficient to induce immunity. Using a protein ligation method, we develop a general approach for linking the immune stimulant, poly dA:dT (pdA:dT), to a monoclonal antibody (mAb) specific for DEC205 (DEC). We show that DEC-specific mAbs deliver pdA:dT…

    Targeted delivery of antigens to dendritic cells (DCs) is a promising vaccination strategy. However, to ensure immunity, the approach depends on coadministration of an adjuvant. Here we ask whether targeting of both adjuvant and antigen to DCs is sufficient to induce immunity. Using a protein ligation method, we develop a general approach for linking the immune stimulant, poly dA:dT (pdA:dT), to a monoclonal antibody (mAb) specific for DEC205 (DEC). We show that DEC-specific mAbs deliver pdA:dT to DCs for the efficient production of type I interferon in human monocyte-derived DCs and in mice. Notably, adaptive T-cell immunity is elicited when mAbs specific for DEC-pdA:dT are used as the activation stimuli and are administered together with a DC-targeted antigen. Collectively, our studies indicate that DCs can integrate innate and adaptive immunity in vivo and suggest that dual delivery of antigen and adjuvant to DCs might be an efficient approach to vaccine development.

    Other authors
    • Scott Barbuto
    • Juliana Idoyaga
    • Paula M Longhi
    • Gael Breton
    • Ralph M. Steinman
    • Tom W. Muir
    See publication

  • Streamlined expressed protein ligation using split inteins

    Journal of the American Chemical Society

    Chemically modified proteins are invaluable tools for studying the molecular details of biological processes, and they also hold great potential as new therapeutic agents. Several methods have been developed for the site-specific modification of proteins, one of the most widely used being expressed protein ligation (EPL) in which a recombinant α-thioester is ligated to an N-terminal Cys-containing peptide. Despite the widespread use of EPL, the generation and isolation of the required…

    Chemically modified proteins are invaluable tools for studying the molecular details of biological processes, and they also hold great potential as new therapeutic agents. Several methods have been developed for the site-specific modification of proteins, one of the most widely used being expressed protein ligation (EPL) in which a recombinant α-thioester is ligated to an N-terminal Cys-containing peptide. Despite the widespread use of EPL, the generation and isolation of the required recombinant protein α-thioesters remain challenging. We describe here a new method for the preparation and purification of recombinant protein α-thioesters using engineered versions of naturally split DnaE inteins. This family of autoprocessing enzymes is closely related to the inteins currently used for protein α-thioester generation, but they feature faster kinetics and are split into two inactive polypeptides that need to associate to become active. Taking advantage of the strong affinity between the two split intein fragments, we devised a streamlined procedure for the purification and generation of protein α-thioesters from cell lysates and applied this strategy for the semisynthesis of a variety of proteins including an acetylated histone and a site-specifically modified monoclonal antibody.

    Other authors
    See publication

  • A Minimalist Design Approach to Antimicrobial Agents Based on a Thionin Template

    Journal of Medicinal Chemistry, 2006, Vol. 49, No. 2 Published on Web 12/24/2005

    Abstract: Numerous studies have been devoted to the stabilization of
    secondary structure elements to improve receptor-ligand recognition.
    We report a novel application of this principle to create new antimicrobial
    agents using the highly folded thionin from Pyrularia pubera
    as a template. Non-native disulfide bonds have been used to induce
    two short linear segments of the protein into an amphipathic helix.
    The resulting 13- and 9-residue peptides are significantly more…

    Abstract: Numerous studies have been devoted to the stabilization of
    secondary structure elements to improve receptor-ligand recognition.
    We report a novel application of this principle to create new antimicrobial
    agents using the highly folded thionin from Pyrularia pubera
    as a template. Non-native disulfide bonds have been used to induce
    two short linear segments of the protein into an amphipathic helix.
    The resulting 13- and 9-residue peptides are significantly more active
    than their linear counterparts and have an activity similar to that of
    native thionin.

    Other authors
    • David Andreu

Languages

  • English

    Native or bilingual proficiency

  • Spanish

    Native or bilingual proficiency

  • Catalan

    Native or bilingual proficiency

More activity by Miquel

View Miquel’s full profile

  • See who you know in common
  • Get introduced
  • Contact Miquel directly
Join to view full profile

Other similar profiles

Explore more posts

Explore collaborative articles

We’re unlocking community knowledge in a new way. Experts add insights directly into each article, started with the help of AI.

Explore More

Add new skills with these courses

See all courses