Expression by Chlamydomonas reinhardtii of a chloroplast ATP synthase with polyhistidine-tagged beta subunits
| Autor: | Richard E. McCarty, Julian N. Rosenberg, Eric A. Johnson |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
0106 biological sciences
Chloroplasts Protein subunit Protozoan Proteins Biophysics Chlamydomonas reinhardtii Polyhistidine Photosynthesis 01 natural sciences Biochemistry 03 medical and health sciences Adenosine Triphosphate Animals Histidine DNA Primers 030304 developmental biology chemistry.chemical_classification CF1 0303 health sciences biology ATP synthase Cell Biology Mitochondrial Proton-Translocating ATPases biology.organism_classification Recombinant Proteins Amino acid Kinetics Protein Subunits Transformation (genetics) Enzyme chemistry biology.protein His-tag 010606 plant biology & botany |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Bioenergetics. 1767(5):374-380 |
| ISSN: | 0005-2728 |
| DOI: | 10.1016/j.bbabio.2007.03.003 |
| Popis: | The green alga Chlamydomonas reinhardtii is a model organism for the study of photosynthesis. The chloroplast ATP synthase is responsible for the synthesis of ATP during photosynthesis. Using genetic engineering and biolistic transformation, a string of eight histidine residues has been inserted into the amino-terminal end of the β subunit of this enzyme in C. reinhardtii. The incorporation of these amino acids did not impact the function of the ATP synthase either in vivo or in vitro and the resulting strain of C. reinhardtii showed normal growth. The addition of these amino acids can be seen through altered gel mobility of the β subunit and the binding of a polyhistidine-specific dye to the subunit. The purified histagged CF1 has normal Mg 2+ -ATPase activity, which can be stimulated by alcohol and detergents and the enzyme remains active while bound to a nickel-coated surface. Potential uses for this tagged enzyme as a biochemical tool are discussed. © 2007 Elsevier B.V. All rights reserved. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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