Using offset recombinant polymerase chain reaction to identify functional determinants in a common family of bacterial albumin binding domains

Biochemistry
David A RozakPhilip Bryan

Abstract

The 46 amino acid GA albumin binding module is a putative virulence factor that has been identified in 16 domains from four bacterial species. Aside from their possible effects on pathogenicity and host specificity, the natural genotypic and phenotypic variations that exist among members of this module offer unique opportunities for researchers to identify and explore functional determinants within the well-defined sequence space. We used a recently developed in vitro recombination technique, known as offset recombinant PCR, to shuffle seven homologues that encode a broad range of natural GA polymorphisms. Phage display and selection were applied to probe the recombinant library for members that showed simultaneous improvements to human and guinea pig serum albumin binding. Thermodynamic data for the most common phage-selected mutant suggest that domain-stabilizing mutations substantially improved GA binding for both species of albumin.

References

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Citations

Sep 18, 2008·Proceedings of the National Academy of Sciences of the United States of America·Yanan HeJohn Orban
May 24, 2008·Protein Engineering, Design & Selection : PEDS·Andreas JonssonPer-Ake Nygren
May 16, 2012·The Journal of Chemical Physics·Ludovico Sutto, Carlo Camilloni
Jul 5, 2007·Proceedings of the National Academy of Sciences of the United States of America·Patrick A AlexanderPhilip N Bryan
Nov 20, 2009·Proceedings of the National Academy of Sciences of the United States of America·Patrick A AlexanderPhilip N Bryan
Jan 18, 2008·Journal of Molecular Recognition : JMR·Oksana Okhrimenko, Ilian Jelesarov
Aug 16, 2015·Protein Engineering, Design & Selection : PEDS·Steven A JacobsKaryn T O'Neil
Jan 1, 2013·Computational and Structural Biotechnology Journal·Johan Nilvebrant, Sophia Hober
Aug 3, 2016·Journal of Molecular Recognition : JMR·Chunnian SongGuangju Chen
Mar 7, 2021·Communications Biology·Yingwei ChenPhilip N Bryan

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