A Cost-effective Amino-acid-type Selective Isotope Labeling of Proteins Expressed in Leishmania tarentolae

0334906 - BC 2010 RIV US eng J - Journal Article
Foldynová-Trantírková, Silvie - Matulová, J. - Dötsch, V. - Löhr, F. - Cirstea, I. - Alexandov, K. - Breitling, R. - Lukeš, Julius - Trantírek, Lukáš
A Cost-effective Amino-acid-type Selective Isotope Labeling of Proteins Expressed in Leishmania tarentolae.
Journal of Biomolecular Structure & Dynamics. Roč. 26, č. 6 (2009), s. 755-761. ISSN 0739-1102
R&D Projects: GA ČR GP204/08/P585; GA AV ČR 1QS600220554; GA AV ČR KAN200100801; GA MŠMT 2B06129
Institutional research plan: CEZ:AV0Z60220518
Keywords : NMR * isotope labeling * protein expression * Leishmania * low-level enrichment
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 1.124, year: 2009

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.
Permanent Link: http://hdl.handle.net/11104/0179515