Engineering of chicken avidin: a progressive series of reduced charge mutants
| Autor: | Meir Wilchek, Markku S. Kulomaa, Olli H. Laitinen, Kari J. Airenne, Tikva Kulik, Ari T. Marttila, Edward A. Bayer |
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| Rok vydání: | 1999 |
| Předmět: |
Streptavidin
DNA Complementary Hot Temperature Mutant Biophysics Biotin Sequence alignment Biology Spodoptera Protein Engineering Biochemistry chemistry.chemical_compound stomatognathic system Structural Biology Genetics Animals Molecular Biology Charge mutant Avidin-biotin technology Rational design Cell Biology Protein engineering respiratory system Avidin DNA-Binding Proteins chemistry Biotinylation biology.protein Mutagenesis Site-Directed Chickens |
| Zdroj: | FEBS letters. 441(2) |
| ISSN: | 0014-5793 |
| Popis: | Avidin, a positively charged egg-white glycoprotein, is a widely used tool in biotechnological applications because of its ability to bind biotin strongly. The high pI of avidin (approximately 10.5), however, is a hindrance in certain applications due to non-specific (charge-related) binding. Here we report a construction of a series of avidin charge mutants with pIs ranging from 9.4 to 4.7. Rational design of the avidin mutants was based on known crystallographic data together with comparative sequence alignment of avidin, streptavidin and a set of avidin-related genes which occur in the chicken genome. All charge mutants retained the ability to bind biotin tightly according to optical biosensor interaction analysis. In most cases, their thermal stability characteristics were indistinguishable from those of the wild-type avidin. Our results demonstrate that the charge properties of avidin can be modified without disturbing the crucial biotin-binding activity. |
| Databáze: | OpenAIRE |
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