HMWGAMMA · Structural characterization of the high molecular weight complex of gamma-secretase
6РП — Действия „Мария Кюри“
- Период
- 2006-04-01 → 2008-03-31
- Финансиране от ЕС
- 145 154 €
- Участници
- 1
- Схема
- IIF
Линиите свързват координатора с партньорите.
Накратко на български
Гама-секретазата и нейните взаимодействия с други протеини в клетъчната мембрана определят как се разграждат определени молекули в мозъка. Разбирането на този процес помага за откриването на нови мишени за лекарства при болестта Алцхаймер.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Final Activity Report Summary - HMWGAMMA (Structural characterization of the high molecular weight complex of gamma-secretase)
Gamma-secretase, an aspartyl protease of the intramembrane cleaving proteases (iCLiPs) family, is responsible for the generation of beta-amyloid peptides (ABETA), which are the primary component of the senile plaques in the brains of Alzheimer disease (AD) patients. Gamma-secretase is a complex containing presenilin, nicastrin, Aph-1 and Pen-2 that cleaves various type I membrane proteins, such as the amyloid precursor protein (APP) and Notch. While these four components are necessary and sufficient for gamma-secretase activity, additional proteins might be involved in its regulation. We established a methodology to purify an active gamma-secretase complex from reconstituted presenilin deficient fibroblasts using tandem affinity purification (TAP) method. We biochemically identified proteins that were associated with active gamma-secretase, including the ones involved in complex maturation, membrane trafficking and the tetraspanin web. An elaborated functional analysis strengthened the hypothesis that various proteins involved in sub-cellular trafficking or complex assembly affected ABETA production. We further corroborated the novel association of gamma-secretase with the tetraspanin web in the cell membrane using co-precipitation and floating assays. In tetraspanin deficient cells several gamma-secretase substrates accumulated and the ratio of long to short ABETA generated by tetraspanin web-associated gamma-secretase was affected. Thus, association with tetraspanin domains tuned gamma-secretase proteolytic activity. Our data provided a first proteome of the gamma-secretase complex and, potentially, novel targets for medication for AD.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
Deposition of the amyloid beta peptide (Abeta) in the brain is critical to the pathogenesis of Alzheimer's Disease (AD). Gamma-secretase is a key enzyme responsible for generating Abeta from its precursor protein.Gamma-secretase has attracted a lot of attention recently because:1) it is a highly unusual protease2) it cleaves many important membrane proteins (Notch, cadherins) and regulates subsequent signalling and3) the enzyme is considered to be a drug target in AD research. Gamma-secretase is the high molecular weight complex composed of at least four essential, integral membrane proteins including presenilin, Nicastrin, Aph-1 and Pen-2.It is particularly important to study the structure-function relationship of gamma-secretase from biological, enzymatical and pathological perspectives. Direct analysis of the structure is difficult because handling such hydrophobic complex in cell and membrane free conditions is extremely problematic. In this project, we plan to take advantage of the camel antibody as a tool for the structural analysis of gamma-secretase. Camel antibodies consist of only heavy chains with highly stable variable regions.The variable region fragment (called nanobody) can be easily expressed as a recombinant protein maintaining specificity and stability. We will immunize camel against the gamma-secretase compex, generate a phage display library of nanobodies, and then screen gamma-secretase binders/inhibitors. Analyses of the obtained inhibitors will provide information about the import ant domains of the gamma-secretase complex. The inhibitors can be applied in a multitude of applications. We will focus on the structural analysis of gamma-secretase, and on cell biological studies. Since these nanobodies can be engineered with modifications that allow them to cross the blood brain barrier, a longer term goal is to use the nanobodies as therapeutics to inhibit gamma-secretase in Alzheimer's Disease.
Оригинален текст от CORDIS (на английски).
Участници
- FLANDERS INTERUNIVERSITY INSTITUTE FOR BIOTECHNOLOGY VZW · ZWIJNAARDEКоординаторБелгия
Връзки
Данни: CORDIS, © Европейски съюз