GLYCOENGINEERING · Enzymatic engineering of modified Glycostructures
6РП — Действия „Мария Кюри“
- Период
- 2006-11-01 → 2007-10-31
- Финансиране от ЕС
- 178 433 €
- Участници
- 1
- Схема
- EIF
Линиите свързват координатора с партньорите.
Накратко на български
Ензимите, които изграждат сложни захарни структури, се модифицират чрез молекулярно моделиране, за да се промени тяхната специфичност, като например при синтеза на кръвни групи А и В. Това помага за по-доброто разбиране на начина, по който ензимите разпознават и свързват своите молекули.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Final Activity Report Summary - GLYCOENGINEERING (Enzymatic Engineering of Modified Glycostructures)
Dr Astrid Blume received training in molecular modelling, protein crystallisation, X-ray crystallography, data refinement and kinetic analysis of two-substrate enzyme systems as outlined in the GlycoEngineering proposal. Molecular modelling included a training course with the MOE program suite. This was subsequently used to design and prepare glycosyltransferase mutants with altered substrate specificities. The X-ray crystallography studies of mutant enzymes were conducted in the laboratory of Professor Stephen Evans at the University of Victoria, Canada. Dr Blume furthered her training in generating mutant enzymes, with the production of blood group A and B synthesising mutant enzymes, GTA and GTB. Additionally she studied the crosstalk between donor and acceptor binding to GTB by STD NMR and evaluated product and substrate binding to the GTA enzyme.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
Oligosaccharides are a diverse class of carbohydrates widely distributed in Nature. Their biological significances range from roles as structural elements and energy sources to being ubiquitous constituents on mammalian, bacterial and viral cell surfaces, where they are involved in cell recognition events.Both the selective modification of cell surface glycan structures and the production of competitive glycan ligands are important research areas with potential for developing novel therapeutics. The chemic al synthesis of oligosaccharides is a multi-step endeavour and is particularly difficult in analog synthesis, where additional steps are required to introduce modifications.Increasingly chemists are employing enzymes to take advantage of their rate acceleration, stereoselectivity, regiospecificity and environmentally friendly green chemistry conditions. Our biocatalytic GlycoEngineering approach uses glycosyltransferases that catalyse the transfer of a monosaccharide from a nucleotide donor to an acceptor.We have identified the blood group A and B synthesizing enzymes (GTA and GTB) as models for studying retaining glycosyltransferases. GTA and GTB will be employed for the synthesis of modified blood group glycostructures, using both enzyme engineering and substrate engineering.Based on X-ray and NMR studies of ligand binding to GTA and GTB, different UDP-N-acetylgalactosamine and UDP-galactose donor analogs will be designed with docking and molecular modelling. To overcome potential limitations of native G TA and GTB in analog preparation, their substrate specificities will be broadened by mutagenesis based on sequence, structure and modelling studies.Donor substrate analogs will be chemi-enzymatically prepared and used with native and mutated GTA and GTB t o synthesize modified oligosaccharide structures. This unique GlycoEngineering approach encompasses engineering both the enzymes and their substrates and can be broadly applied to other glycosyltransferases.
Оригинален текст от CORDIS (на английски).
Участници
- CARLSBERG LABORATORY · VALBYКоординаторДания
Връзки
Данни: CORDIS, © Европейски съюз