Cost-effective method for the preparation of uniformly labeled myristoylated proteins for NMR measurements

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60461373%3A22330%2F14%3A43897513" target="_blank" >RIV/60461373:22330/14:43897513 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/60461373:22810/14:43897513

Výsledek na webu

<a href="http://www.sciencedirect.com/science/article/pii/S1046592814000552#" target="_blank" >http://www.sciencedirect.com/science/article/pii/S1046592814000552#</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.pep.2014.03.005" target="_blank" >10.1016/j.pep.2014.03.005</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Cost-effective method for the preparation of uniformly labeled myristoylated proteins for NMR measurements

Popis výsledku v původním jazyce

Nuclear magnetic resonance (NMR) is a powerful technique for solving protein structures or studying their interactions. However, it requires molecules labeled with NMR sensitive isotopes like carbon C-13 and nitrogen N-15. The recombinant expression of labeled proteins is simple to perform but requires quite expensive chemicals. When there is a need for special labeled chemicals, like uniformly C-13-labeled myristic acid, the price significantly rises. Here we describe a cost-effective method for the recombinant expression of uniformly labeled myristoylated proteins in Escherichia coli demonstrated on the production of Mason-Pfizer monkey virus matrix protein. We used the ability of E. coli to naturally synthetize myristic acid. When grown in isotopically labeled medium the myristic acid will be labelled as well. Bacteria were co-transfected with plasmid carrying gene for yeast N-myristoyltransferase which ensures myristoylation of expressed protein. This process provided 1.8 mg of the

Název v anglickém jazyce

Cost-effective method for the preparation of uniformly labeled myristoylated proteins for NMR measurements

Popis výsledku anglicky

Nuclear magnetic resonance (NMR) is a powerful technique for solving protein structures or studying their interactions. However, it requires molecules labeled with NMR sensitive isotopes like carbon C-13 and nitrogen N-15. The recombinant expression of labeled proteins is simple to perform but requires quite expensive chemicals. When there is a need for special labeled chemicals, like uniformly C-13-labeled myristic acid, the price significantly rises. Here we describe a cost-effective method for the recombinant expression of uniformly labeled myristoylated proteins in Escherichia coli demonstrated on the production of Mason-Pfizer monkey virus matrix protein. We used the ability of E. coli to naturally synthetize myristic acid. When grown in isotopically labeled medium the myristic acid will be labelled as well. Bacteria were co-transfected with plasmid carrying gene for yeast N-myristoyltransferase which ensures myristoylation of expressed protein. This process provided 1.8 mg of the

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CE - Biochemie

OECD FORD obor

—

Projekt

<a href="/cs/project/GAP302%2F12%2F1895" target="_blank" >GAP302/12/1895: Role retrovirového matrixového proteinu při transportu virové částice a její interakci s cytoplasmatickou membránou.</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2014

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Protein Expression and Purification

ISSN

1046-5928

e-ISSN

—

Svazek periodika

99

Číslo periodika v rámci svazku

červenec 2014

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

4

Strana od-do

6-9

Kód UT WoS článku

000336870800002

EID výsledku v databázi Scopus

—