ARRESTED RHODOPSIN · Crystallisation and functional analysis of rhodopsin bound to its effector arrestin
6РП — Действия „Мария Кюри“
- Период
- 2007-01-01 → 2008-12-31
- Финансиране от ЕС
- 159 046 €
- Участници
- 1
- Схема
- EIF
Линиите свързват координатора с партньорите.
Накратко на български
Родопсинът в човешкото око се използва, за да се разбере как протеините пренасят сигнали през клетъчната мембрана. Подоброто познаване на тези процеси помага при разработването на нови лекарства за регулиране на сърдечния ритъм, храносмилането или невронната активност.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Final Activity Report Summary - ARRESTED RHODOPSIN (Crystallisation and functional analysis of rhodopsin bound to its effector arrestin)
The role of G protein-coupled receptors (GPCRs) in humans is of great medical importance. Since these proteins can interact with the environment both inside and outside the cell, they act as a receptor of chemical signals that activate a variety of cellular reactions. Many medical drugs mimic these signals to influence body reactions like heart beat, digestion or neuronal activity. The development of future drugs is facilitated by a detailed understanding of how GPCRs transfer their signal across the cellular membrane. A crucial key for our understanding is the availability of high-resolution structures illuminating all steps of the GPCR reaction cycle. Rhodopsin, the dim light sensor of the human eye, gives us the unique opportunity to follow these reaction cycles. Consequently, rhodopsin is the only GPCR for which structures of several intermediates were already determined, much of which was done in the laboratory which hosted the fellow. The Marie Curie funding allowed for significant progress towards visualising the fully active conformation of rhodopsin. To do this, human cells were altered to produce genetically modified rhodopsin in a stable form, i.e. a form that could be used more easily by the techniques applied to determine the structure of proteins. The structure of this stabilised rhodopsin could therefore be solved, and this was the first time that this was possible for a GPCR produced by recombinant technology. Several additional alterations to the receptor were also created to allow for the stable binding of arrestin and G protein, proteins known to arrest GPCRs in their active conformation. Our next challenge would be to determine the structure of such an activated receptor complex so as to find out how signals were transmitted. Our knowledge of these receptor structures would allow us to make significant steps towards a better treatment of GPRC-related illnesses.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
Transmission of signals across the plasma membrane is among the most fundamental cellular processes and is medically highly relevant. The largest group of membrane proteins involved in this process are the G protein coupled receptors (GPCRs). Despite their broad physiological relevance, GPCRs share a seven-a-helix architecture and transmit the activation signal by a heterotrimeric guanyl nucleotide-binding protein (G-protein). Also desensitisation of GPCRs occurs via highly conserved mechanisms that involve phosphorylation of the receptor and binding of a protein class called arrestins. Rhodopsin, the photoreceptor protein in retina rod cells, is a prototypical GPCR. My host laboratory has determined structures of rhodopsin in the ground and the metarhodopsin I intermediate state using x-ray and electron crystallography, respectively.The next logical step is to solve the structure of fully activated rhodopsin in complex with its effectors. The arrestin/rhodopsin complex is the most promising target for such a purpose since its classical function is to down regulate signal transduction for longer times and thus the complex is expected to be stable. My host laboratory and their international co-operators are able to purify all components known to be involved in arresting rhodopsin in mg amounts either from native tissue or recombinant sources. Several constitutively activated mutants of rhodopsin will be coupled with tagged arrestin on Ni-chelating and ConA columns. New fluorescence assays developed by our co-operators and at the MRC-LMB will allow rapid determination of conditions optimal for complex formation.Once arrestin/rhodopsin complexes are formed, they will be biochemically characterised and crystallised using state of the art robot facilities. Structure determination will be carried out with molecular replacement revealing the fully activated receptor and its interacting surface with arrestin in molecular detail.
Оригинален текст от CORDIS (на английски).
Участници
- MEDICAL RESEARCH COUNCIL · LONDONКоординаторОбединеното кралство
Връзки
Данни: CORDIS, © Европейски съюз