Directed evolution to probe protein allostery and integrin I domains of 200,000-fold higher affinity
| العنوان: | Directed evolution to probe protein allostery and integrin I domains of 200,000-fold higher affinity |
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| المؤلفون: | Moonsoo M. Jin, Motomu Shimaoka, Yong Sung Kim, Nathan S. Astrof, Dane K. Wittrup, Timothy A. Springer, Christopher V. Carman, Gang Song |
| المصدر: | Proceedings of the National Academy of Sciences. 103:5758-5763 |
| بيانات النشر: | Proceedings of the National Academy of Sciences, 2006. |
| سنة النشر: | 2006 |
| مصطلحات موضوعية: | Models, Molecular, Integrins, Saccharomyces cerevisiae Proteins, Molecular Sequence Data, Allosteric regulation, Intercellular Adhesion Molecule-1, Integrin, Mutant, Biology, Polymerase Chain Reaction, Protein Structure, Secondary, Protein structure, Allosteric Regulation, Amino Acid Sequence, Binding site, Binding Sites, Multidisciplinary, Proteins, Biological Sciences, Surface Plasmon Resonance, Ligand (biochemistry), Directed evolution, Recombinant Proteins, Cell biology, Kinetics, Solubility, biology.protein, Directed Molecular Evolution |
| الوصف: | Understanding allostery may serve to both elucidate mechanisms of protein regulation and provide a basis for engineering active mutants. Herein we describe directed evolution applied to the integrin αLinserted domain for studying allostery by using a yeast surface display system. Many hot spots for activation are identified, and some single mutants exhibit remarkable increases of 10,000-fold in affinity for a physiological ligand, intercellular adhesion molecule-1. The location of activating mutations traces out an allosteric interface in the interior of the inserted domain that connects the ligand binding site to the α7-helix, which communicates allostery to neighboring domains in intact integrins. The combination of two activating mutations (F265S/F292G) leads to an increase of 200,000-fold in affinity to intercellular adhesion molecule-1. The F265S/F292G mutant is potent in antagonizing lymphocyte function-associated antigen 1-dependent lymphocyte adhesion, aggregation, and transmigration. |
| تدمد: | 1091-6490 0027-8424 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::83d8c8c2a871ce540e47dfb7c1fff00f https://doi.org/10.1073/pnas.0601164103 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....83d8c8c2a871ce540e47dfb7c1fff00f |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10916490 00278424 |
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