H2020Индивидуална стипендия2015–2017

ACPNMR · Structural dynamics of acyl carrier protein complexes through combined solution and solid-state NMR

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

Период: 2015-09-10 → 2017-09-09
Финансиране от ЕС: 183 455 €
Участници: 1
Схема: MSCA-IF-EF-ST

Линиите свързват координатора с партньорите.

Накратко на български

Структурната динамика на протеинови комплекси, участващи например в прикрепянето на хлор при синтеза на природни съединения, се анализира чрез ЯМР спектроскопия. Познаването на тези процеси помага за по-ефективното създаване на модифицирани ензими за производство на лекарства срещу бактериални патогени.

Този кратък обзор е генериран от изкуствен интелект

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

Резултати накратко

Structural dynamics of acyl carrier protein complexes through combined solution and solid-state NMR

The aim of the proposed project was to obtain molecular level understanding of protein interactions in complexes from biosynthetic factories for natural products active against bacterial pathogens. Natural products including polyketides and non-ribosomal peptides comprise 20% of the top-selling drugs, with combined worldwide annual revenues of over £10 billion. Due their complex structures, they are often difficult to synthesize by chemical methods. However, a synthetic biology approach where the natural products are produced biosynthetically by genetically manipulated assembly lines, which include non-ribosomal peptide synthases (NRPSs) and polyketide syntheses (PKSs), is becoming increasingly popular. Inactivation, substitution or addition of new domains to the naturally occurring modules, enables the biosynthesis of compounds that are not found in nature and introduce desirable properties into natural products. However, engineering of NRPSs/PKSs often leads to assembly lines that are less efficient than the original multienzymes or sometimes not active. Knowledge of the structural dynamics exhibited by the NRPSs is therefore essential to enable their engineering, especially if such strategies are to be ever implemented in an industrial scale production. As a general approach we proposed to use a combined solution and solid-state NMR to study such systems. In particular, in this project we have investigated systems involved in important biosynthetic step of halogenation (specifically attachment of chlorine). Using multidisciplinary approach including biochemical assays, biophysical methods, NMR, mass spectrometry and X-ray crystallography we obtained initial insight for the mechanism controlling the concerned reaction.

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

Цел на проекта

Antibiotics are a vital part of modern medicine. However, the available arsenal of antibiotics becomes less effective as microorganisms develop ""resistance"" against them. The resulting crisis in medicine necessitates development of new drugs. Natural products inspired compounds are a potential solution to this challenge. For example, gladiolin biosythesized by a mulitenzyme polyketide synthase (PKS) was shown to be active against Mycobacterium tuberculosis, a multidrug resistant bacterium that one third of world’s population is infected with. The PKS producing gladiolinum is a good example of multienzymatic assembly lines that due to their modular nature are ideal for genetic manipulation paving the way for synthetic biology approach to produce new drugs (that are difficult to synthesize using chemical methods). However, for such approach to be successful it is crucial to understand molecular level structural and dynamical factors responsible for controlling directionality and specificity of biosynthesis. Neglecting such factors, when modifying PKSs often results in assembly lines that are inactive or dysfunctional. Here we propose to use a novel approach combining state-of-the-art solution and solid-state NMR methods to investigate structure, dynamics and interactions of proteins from module 12 of gladiolin PKS, particularly acyl carrier proteins (ACP12a and ACP12b) and special adapter ketosynthase (KS12), all of them highly required in industrial biosynthesis toolbox. We will use solution NMR to characterize isolated ACPs and solid-state NMR to study ACPs-KS12 complexes (direct structural information is difficult to obtain by solution NMR due to the large complex size). Combining solution and solid-state NMR relaxation methods will allow us to probe protein motions over 6 orders of magnitude providing a comprehensive picture of relevant dynamic changes in ACPs-KS12 complexes.""

Оригинален текст от CORDIS (на английски).

Участници

UNIVERSITY OF WARWICK · COVENTRYКоординаторОбединеното кралство