H2020Индивидуална стипендия2016–2019

DIAL-IN · Dynamic Integrated structural and proteomic Analysis of the LUBAC complex and its involvement in NF-κB and inflammation

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

Период: 2016-11-01 → 2019-03-02
Финансиране от ЕС: 175 420 €
Участници: 1
Схема: MSCA-IF-EF-ST

Линиите свързват координатора с партньорите.

Накратко на български

Протеиновият комплекс LUBAC и неговата роля в сигналите за възпаление се анализират чрез масспектрометрия. Разбирането на тези процеси помага да се разберат механизмите на възпалението, рака и стареенето.

Този кратък обзор е генериран от изкуствен интелект

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

Резултати накратко

Dynamic Integrated structural and proteomic Analysis of the LUBAC complex and its involvement in NF-κB and inflammation

PROBLEM AND RELEVANCE: TNF-α is among the most critical molecules regulating the inflammatory response in mammalians. By binding to its cognate receptor, TNF-RI, the pleiotropic cytokine TNF-α induces the formation of a large multiprotein complex (the TNF-RSC - tumor necrosis factor receptor signaling complex), which relays the signal to several transcription factors and eventually regulates the response of the cell to injury and pathogens invasion. Depending on specific conditions, the signaling can be rerouted to promote different type of cell deaths - apoptosis and necroptosis. Given its key role in live-or-die cell fate, TNF-α plays not only a prominent role in inflammation, but also in cancer and ageing. While an exhaustive understanding of the TNF-RSC would be instrumental to control and interpret the outcomes of the TNF signaling, and in spite of decades of studies, we are still far from achieving this goal. Among the many poorly or incompletely understood aspects, several can be mentioned, including (i) the architecture and stoichiometry of the complex; (ii) a complete catalogue of the members; (iii) regulation of its progression by phosphatases; (iv) dynamics of internalisation; (v) the organisation at the membrane. OBJECTIVES: As part of my Marie Curie project, I proposed to characterize the TNF-RSC by a variety of complementary methods, based mostly on mass spectrometry, to obtain a comprehensive understanding of the composition, dynamics and regulation of this most relevant complex. A particular emphasis has been placed on the ubiquitin ligase LUBAC, a recently discovered protein complex that is part of the TNF-RSC and that has not yet been, in many respect, thoroughly characterised.

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

Цел на проекта

One of key challenges in current biology is to characterize the genotype to phenotype transition and explain how genomic variation results in specific phenotypes. According to the 'modular biology' paradigm, dynamically regulated multi-protein modules mediate this transition. My project aims at testing this paradigm by applying to a clinically highly relevant case study an integrated workflow of new technologies developed in the host laboratory. In particular, I will focus on a novel ubiquitin ligase complex, linear ubiquitin chain assembly complex (LUBAC), a critical but poorly understood player in several inflammation-related pathways, most notably the canonical NF-κB pathway. Given its key role in the pathogenesis of inflammation, understanding LUBAC will have fundamental biomedical implications. To investigate its structure, function and regulation, I will integrate data generated by mass spectrometry, which is the focus of the Aebersold laboratory, and cryo-electron microscopy (cryo-EM), which is the main expertise I acquired during my PhD. First, I will use quantitative affinity-purification coupled to mass spectrometry (AP-SWATH) to identify the dynamic interactome and the post-translation modifications (PTMs) of LUBAC. I will then define the structure of the complex by combining available atomic models of known LUBAC components, cryo-EM and cross-linking coupled to mass spectrometry (XL-MS). Finally, I will study clinical mutants and map the repercussions of genetic variation at the level of the complex organization, its interactome and of the global proteomic landscape. By these means, I will give a systemic and mechanistic account of the genotype to phenotype transition in clinically relevant scenarios. The output of the project, in line with the Work Programme, will be the production of new knowledge of fundamental biological and medical relevance and the development and validation of enabling technologies at the forefront of molecular biology research.

Оригинален текст от CORDIS (на английски).

Участници

EIDGENOESSISCHE TECHNISCHE HOCHSCHULE ZUERICH · ZuerichКоординаторШвейцария