Structural basis for function within the family of nitric oxide synthases
Project goals
The family of nitric oxide synthases (inducible-iNOS; endothelial-eNOS; neuronal-nNOS) combines the unique properties of heme- and flavin-containing proteins in one polypeptide chain to achieve the, production of a small, diffusible, reactive biologicalmessenger and cytotoxic agent, nitric oxide (NO). NO functions in cell-to-cell signaling and plays a critical role in numerous physiological processes. The aberrant production of NO is responsible for a number of pathologies. The crystal structures of eNOS, iNOS, and nNOS heme domain and that of FAD/NADPH domain of nNOS have been solved. NOS is an important drug target and the structural details of holoenzyme are essential for this. However, a complete structure is unavailable either for the reductasedomain or the holoenzyme. To accomplish this goal we will utilize mammalian and prokaryotic expression systems to overexpress (i) all three NOS isoforms and (ii) its naturally occurring variants (muscle specific variant, ĽNOS, iNOS bronchial epithelium
Keywords
Public support
Provider
Czech Science Foundation
Programme
Standard projects
Call for proposals
Standardní projekty 8 (SGA02005GA-ST)
Main participants
—
Contest type
VS - Public tender
Contract ID
303/05/0336
Alternative language
Project name in Czech
Strukturální základy funkce rodiny syntáz oxidu dusnatého
Annotation in Czech
Syntázy oxidu dusnatého (tři savčí isoformy: indukovatelna forma-iNOS; endotelová-eNOS; neuronální-nNOS) jsou neobvyklé hemoflavoproteiny. Výsledkem jejich enzymatické aktivity je tvorba oxidu dusnatého (NO). Tato malá, difuzibilní a reaktivní molekula hraje důležitou roli v mezibuněčné signalizaci a účastní se řady klíčových fyziologických pochodů. Vychýlená produkce NO hraje důležitou roli v patogeneze aterosklerózy, septického šoku, nádorových onemocnění, neurodegenerace a dalších chorobných stavů. Na ovlivnění aktivity NOS je proto zaměřena řada farmakologických intervencí. Poznání kompletní struktury NOS je důležitým předpokladem cíleného farmakologického zásahu. Byly určeny krystalové struktury hemových domén iNOS, eNOS, nNOS a větší části reduktázové domény nNOS. Přes mnohaleté úsilí nebyla popsána struktura kompletní NOS ani její kompletní reduktázová doména. Chceme pomocí savčích a prokaryotických expresních systémů připravit (i) všechny izoformy NOS a (ii) jejich vybrané varianty [nNOS
Scientific branches
R&D category
ZV - Basic research
CEP classification - main branch
CE - Biochemistry
CEP - secondary branch
EB - Genetics and molecular biology
CEP - another secondary branch
EI - Biotechnology and bionics
10603 - Genetics and heredity (medical genetics to be 3)
10604 - Reproductive biology (medical aspects to be 3)
10605 - Developmental biology
10608 - Biochemistry and molecular biology
10609 - Biochemical research methods
20801 - Environmental biotechnology
20802 - Bioremediation, diagnostic biotechnologies (DNA chips and biosensing devices) in environmental management
20803 - Environmental biotechnology related ethics
20901 - Industrial biotechnology
20902 - Bioprocessing technologies (industrial processes relying on biological agents to drive the process) biocatalysis, fermentation
20903 - Bioproducts (products that are manufactured using biological material as feedstock) biomaterials, bioplastics, biofuels, bioderived bulk and fine chemicals, bio-derived novel materials
30101 - Human genetics
30401 - Health-related biotechnology
30402 - Technologies involving the manipulation of cells, tissues, organs or the whole organism (assisted reproduction)
30403 - Technologies involving identifying the functioning of DNA, proteins and enzymes and how they influence the onset of disease and maintenance of well-being (gene-based diagnostics and therapeutic interventions [pharmacogenomics, gene-based therapeutics])
30404 - Biomaterials (as related to medical implants, devices, sensors)
30405 - Medical biotechnology related ethics
40401 - Agricultural biotechnology and food biotechnology
40402 - GM technology (crops and livestock), livestock cloning, marker assisted selection, diagnostics (DNA chips and biosensing devices for the early/accurate detection of diseases) biomass feedstock production technologies, biopharming
40403 - Agricultural biotechnology related ethics
Completed project evaluation
Provider evaluation
V - Vynikající výsledky projektu (s mezinárodním významem atd.)
Project results evaluation
The family of nitric oxide synthases (NOS) combines together into one protein the unique properties of hemoproteins and flavoproteins. The result of their enzymatic, immensely complex controlled action is the generation of a small, rapidly diffusing bior
Solution timeline
Realization period - beginning
Jan 1, 2005
Realization period - end
Dec 31, 2007
Project status
U - Finished project
Latest support payment
May 2, 2007
Data delivery to CEP
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data delivery code
CEP08-GA0-GA-U/04:3
Data delivery date
Dec 16, 2008
Finance
Total approved costs
3,201 thou. CZK
Public financial support
3,201 thou. CZK
Other public sources
0 thou. CZK
Non public and foreign sources
0 thou. CZK
Recognised costs
3 201 CZK thou.
Public support
3 201 CZK thou.
0%
Provider
Czech Science Foundation
CEP
CE - Biochemistry
Solution period
01. 01. 2005 - 31. 12. 2007