Protein Library Design
Design of protein-binding recognition and specificity
Due to their function and specificity, proteins are increasingly being used as platforms for a wide-range of research, biotechnical, synthetic biology and medical applications. However, challenges associated with introducing a novel function into a protein with evolved physical and functional properties, as well as high stringency of protein-protein interaction specificity necessary to avoid unwanted effects, often hinder these applications. Thus, to take advantage of powerful library approaches towards successfully engineering proteins for downstream biomedical and synthetic biology applications, protein binding and specificity must be included in the library design. We are applying and developing approaches to high-quality protein library design and selections, including post-translational modifications, towards understanding the basis of protein recognition and binding specificity in pervasive-binding model systems.
Selected Publications
Bradley LH. (2014) High-quality combinatorial protein libraries using the binary patterning approach. Methods Mol Biol, 1216:117-28.
Magnani R, Chaffin B, Dick E, Bricken ML, Houtz RL, Bradley LH. (2012) Utilization of a calmodulin lysine methyltransferase co-expression system for the generation of a combinatorial library of post-translationally modified proteins. Protein Expression & Purification, 86: 83-88. http://dx.doi.org/10.1016/j.pep.2012.09.012
Arai R, Kobayashi N, Kimura A, Sato T, Matsuo K, Wang AF, Platt JM, Bradley LH, Hecht MH. (2012) Domain-Swapped Dimeric Structure of a Stable and Functional De Novo 4-Helix Bundle Protein, WA20. J. Phys. Chem. B. 116: 6569-7032. http://pubs.acs.org/doi/abs/10.1021/jp212438h
Sexton T, Hitchcook LJ, Rodgers DW, Bradley LH, Hersh LB (2012) Active Site Mutations Change the Cleavage Specificity of Neprilysin. PLoS ONE 7(2): e32343. http://dx.plos.org/10.1371/journal.pone.0032343
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Bradley LH, Bricken ML, Randle C. (2011) Expression, purification, and characterization of proteins from high-quality combinatorial libraries of the mammalian calmodulin central linker.
Protein Expression & Purification, 75(2):186-192. http://dx.doi.org/10.1016/j.pep.2010.08.007
Fisher MA, McKinley KL, Bradley LH, Viola SR, Hecht MH. (2011) De novo designed proteins from a library of artificial sequences function in Escherichia Coli and enable cell growth. PLoS ONE 6(1): e15364. http://dx.doi.org/10.1371/journal.pone.0015364
Patel SC, Bradley LH, Jinadasa SP, Hecht MH. (2009) Cofactor binding and enzymatic activity in an unevolved superfamily of de novo designed 4-helix bundle proteins. Protein Science, 18:1388-1400. http://dx.doi.org/10.1002/pro.147
Bradley LH, Thumfort PP, Hecht MH. (2006) De novo proteins from binary-patterned combinatorial libraries. Methods Mol Biol. 340: 53-69.
Bradley LH, Kleiner RE, Wang AF, Hecht MH, Wood DW. (2005) An intein-based genetic selection allows the construction of a high-quality library of binary patterned de novo protein sequences. Protein Engineering, Design & Selection 18: 201-207.
Design of protein-binding recognition and specificity
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