H2020Индивидуална стипендия2017–2019

PROFOLIG · Covalent-ligation-assisted elucidation of protein-aromatic foldamer interactions

„Хоризонт 2020“ — Действия „Мария Склодовска-Кюри“

Период: 2017-03-01 → 2019-02-28
Финансиране от ЕС: 185 076 €
Участници: 1
Схема: MSCA-IF-EF-ST

Линиите свързват координатора с партньорите.

Накратко на български

Ароматните фолдамери се тестват като молекули, които разпознават повърхностите на протеини, като например човешката карбонанова анхидраза II. Това помага за разработването на нови терапевтични подходи за блокиране на взаимодействията между протеини, които участват в различни заболявания.

Този кратък обзор е генериран от изкуствен интелект

Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.

Резултати накратко

Covalent-ligation-assisted elucidation of protein-aromatic foldamer interactions

Protein-protein interactions (PPIs) play crucial roles in many biological processes and diseases, and represent potential targets to develop new therapeutic approaches. However, PPI inhibition often requires ligands that can cover large areas of protein surfaces, such as other proteins (eg antibodies). With their medium size, the stability and predictability of their helical folded structures, aromatic foldamers constitute candidates for protein surface recognition. The main objectives of this project consisted in the development of aromatic foldamers to interact specifically with the protein surfaces by anchoring approaches in aqueous media towards the development of specific binding ligands in the absence of anchor. The devised strategy consisted in an iterative design process that exploits the structural information of folded synthetic oligomer sequences interacting with protein surfaces to implement subsequent refinements. Starting with a well-studied protein i.e. human carbonic anhydrase II (HCA), we used an inhibitor strategy as first-principles design to obtain structural information of the interactions between the anchored foldamer and HCA protein surface that considers features such as helical length, proteinogenic side chains and hydrogen bonding ability. This anchoring approach was then extended to a covalent strategy in the case of therapeutically interesting proteins with large contact surfaces involved in protein-protein interactions (PPI) such as cyclophilin A (CypA), interleukin-4 (IL4) and ubiquitin (Ub). We finally envisioned to remove the covalent linker in the case of protein/foldamer complexes with stronger affinity. The methodology achieved the following objectives: a) synthesis of a library of foldamer sequences of different length containing various proteinogenic side chains, and anchoring them to protein surfaces; b) screening of the protein-foldamer interactions by induced circular dichroism (ICD); c) assessing the structural information either by solid state structures (X-ray crystallography) or by solution NMR studies; d) new design optimization based on the structural information.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз

Цел на проекта

The development of ligands that selectively bind to a given protein surface is a challenging area of research with potential applications in diagnostics, pharmacology or therapy. Current approaches include small molecule screening, the design of medium sized epitope mimetics (e.g. alpha helix mimetics) and directed evolution methodologies (e.g. ribosome display).In this project, we intend to initiate an essentially unexplored approach: the design from first principles of synthetic ligands derived from helical aromatic oligoamide foldamer backbones bearing proteinogenic side chains to target the surface of a given protein: interleukin 4 (IL4). Helical aromatic amides appear to be well suited for this purpose thanks to their predictable, tunable and stable conformations in solution; their relatively easy synthesis of secondary and tertiary-like structures as large as small proteins; and their high amenability to crystal growth and structural elucidation. Specifically, we intend to explore molecular recognition rules between large (2-15 kDa) aromatic oligoamide foldamers and a target protein surface, and to validate a novel iterative method based on combining covalent attachment and structural characterization.The proposed strategy thus consists in structure-based iterative design. The following steps will be implemented: 1) synthesis of a small pool of foldamer sequences; 2) covalent attachment of each of them via a disulfide bridge to the surface of a recombinantly expressed IL4 cystein mutant, and screening for foldamer-protein interactions through foldamer helix handedness induction by the chiral protein surface; 3) structural characterization of the interactions, mainly by crystallography, within selected protein-foldamer adducts; 4) design of new and improved foldamers. Ultimately optimized interactions should produce foldamers that bind to IL4 without the assistance of the covalent tether.

Оригинален текст от CORDIS (на английски).

Участници

CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS · ParisКоординаторФранция