Number of the records: 1
A Cost-effective Amino-acid-type Selective Isotope Labeling of Proteins Expressed in Leishmania tarentolae
- 1.
SYSNO ASEP 0334906 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title A Cost-effective Amino-acid-type Selective Isotope Labeling of Proteins Expressed in Leishmania tarentolae Author(s) Foldynová-Trantírková, Silvie (BC-A) RID
Matulová, J. (CZ)
Dötsch, V. (DE)
Löhr, F. (DE)
Cirstea, I. (DE)
Alexandov, K. (DE)
Breitling, R. (DE)
Lukeš, Julius (BC-A) RID, ORCID
Trantírek, Lukáš (BC-A)Source Title Journal of Biomolecular Structure & Dynamics. - : Taylor & Francis - ISSN 0739-1102
Roč. 26, č. 6 (2009), s. 755-761Number of pages 7 s. Language eng - English Country US - United States Keywords NMR ; isotope labeling ; protein expression ; Leishmania ; low-level enrichment Subject RIV EB - Genetics ; Molecular Biology R&D Projects GP204/08/P585 GA ČR - Czech Science Foundation (CSF) 1QS600220554 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR) KAN200100801 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR) 2B06129 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) CEZ AV0Z60220518 - PAU-O, BC-A (2005-2011) UT WOS 000266300700008 Annotation We report a cost efficient approach for amino-acid-type selective isotope labeling of proteins expressed in Leishmania tarentolae. The method provides an economically advantageous alternative to recently established protocol for isotopic labeling using expensive synthetic media. The method is based on cultivation of the L. tarentolae expression strain in a cheap complex medium supplemented with labeled amino acid(s). In this protocol, a labeled amino acid is deliberately diluted in the medium of undefined composition, which leads to alow-level isotope enrichment upon protein over-expression. The economic advantage of the protocol is achieved by avoiding large volumes of expensive synthetic medium. Decreased sensitivity of a NMR experiment due to low-level isotope enrichment is compensated by a five- to seven-fold increase of the yield of the recombinant protein in complex mediumas compared to that in the synthetic medium. Workplace Biology Centre (since 2006) Contact Dana Hypšová, eje@eje.cz, Tel.: 387 775 214 Year of Publishing 2010
Number of the records: 1