دورية أكاديمية
Isotopic labeling of recombinant proteins expressed in the protozoan host Leishmania tarentolae
| العنوان: | Isotopic labeling of recombinant proteins expressed in the protozoan host Leishmania tarentolae |
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| المؤلفون: | Niculae, Anca, Bayer, Peter, Cirstea, Ion, Bergbrede, Tim, Pietrucha, Regina, Gruen, Mathias, Breitling, Reinhard, Alexandrov, Kirill |
| بيانات النشر: | Academic Press |
| سنة النشر: | 2006 |
| المجموعة: | The University of Queensland: UQ eSpace |
| مصطلحات موضوعية: | N-15-labeling, Recombinant protein, Eukaryotic expression system, Green fluorescent protein (GFP), Insect cells, Amino-acids, Purification, System, 0304 Medicinal and Biomolecular Chemistry, 06 Biological Sciences, 0601 Biochemistry and Cell Biology |
| الوصف: | Isotope labeling of recombinant proteins is a prerequisite for application of nuclear magnetic resonance spectroscopy (NMR) for the characterization of the three-dimensional structures and dynamics of proteins. Overexpression of isotopically labeled proteins in bacterial or yeast host organisms has several drawbacks. In this work, we tested whether the recently described eukaryotic protein expression system based on the protozoa Leishmania tarentolae could be used for production of amino acid specific 15N-labeled recombinant proteins. Using synthetic growth medium we were able to express in L. tarentolae and purify to homogeneity (15)N-valine labeled Enchanced Green Fluorescent Protein (EGFP) with the final yield of 5.7 mg/liter of suspension culture. NMR study of isolated EGFP illustrated the success of the labeling procedure allowing identification of all 18 valine residues of the protein in the HSQC spectrum. Our results demonstrate the suitability of the L. tarentolae expression system for production of isotopically labeled proteins. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| تدمد: | 1046-5928 1096-0279 |
| العلاقة: | orcid:0000-0002-0957-6511 |
| الإتاحة: | https://doi.org/10.1016/j.pep.2006.04.006Test https://espace.library.uq.edu.au/view/UQ:186517Test |
| رقم الانضمام: | edsbas.10108EB1 |
| قاعدة البيانات: | BASE |
| تدمد: | 10465928 10960279 |
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