Intramolecular cooperativity in a protein binding site assessed by combinatorial shotgun scanning mutagenesis
| Autor: | Kevin P. Clark, Gábor Pál, Sachdev S. Sidhu, Mark Ultsch, Anthony A. Kossiakoff, Bridget Currell |
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| Rok vydání: | 2004 |
| Předmět: |
Models
Molecular Binding Sites Stereochemistry Chemistry Human Growth Hormone Mutagenesis Membrane Proteins Cooperativity Plasma protein binding Protein engineering Affinities Protein Structure Tertiary Protein structure Biochemistry Structural Biology Intramolecular force Humans Binding site Molecular Biology Protein Binding |
| Zdroj: | Journal of molecular biology. 347(3) |
| ISSN: | 0022-2836 |
| Popis: | Combinatorial shotgun alanine-scanning was used to assess intramolecular cooperativity in the high affinity site (site 1) of human growth hormone (hGH) for binding to its receptor. A total of 19 side-chains were analyzed and statistically significant data were obtained for 145 of the 171 side-chain pairs. The analysis revealed that 90% of the side-chain pairs exhibited no statistically significant pair interactions, and the remaining 10% of side-chain pairs exhibited only small interactions corresponding to cooperative interaction energies with magnitudes less than 0.4 kcal/mol. The statistical predictions were tested by measuring affinities for purified mutant proteins and were found to be accurate for five of six side-chain pairs tested. The results reveal that hGH site 1 behaves in a highly additive manner and suggest that shotgun scanning should be useful for assessing cooperative effects in other protein-protein interactions. |
| Databáze: | OpenAIRE |
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