PMID: 11312694Apr 21, 2001Paper

Mechanistic investigation of smart polymer-protein conjugates

Bioconjugate Chemistry
Tsuyoshi ShimobojiAllan S Hoffman

Abstract

Many affinity separation and diagnostic applications rely upon both capture and release steps. There is thus a need for methods to enhance the reversibility of biomolecular interactions. We have previously demonstrated that stimuli-responsive polymers can be used to gate biomolecular reactions when conjugated near the active site of proteins. Here we have used a new smart polymer, N,N-dimethyl acrylamide-co-4-phenylazophenylacrylate that has allowed a mechanistic investigation of the smart polymer switches. This polymer was conjugated via a vinyl sulfone terminus to cysteine residues of genetically engineered streptavidin mutant E116C, where the polymer is conjugated close to the biotin-binding site, and streptavidin mutant S139C, where the conjugation site is distant. The biotin binding switching activity was strongly dependent on conjugation position, as the E116C conjugate displayed a large thermal response while the S139C conjugate displayed only small effects. Kinetic measurements of biotin release demonstrated that the off-rate of biotin was unperturbed and that the thermally triggered release of biotin with the E116C conjugate was due to the blocking the reassociation of biotin. The addition of free polymer to purified E...Continue Reading

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Citations

Dec 18, 2002·Proceedings of the National Academy of Sciences of the United States of America·Tsuyoshi ShimobojiPatrick S Stayton
Oct 25, 2006·Applied Microbiology and Biotechnology·Pall ThordarsonKelly Velonia
Jun 18, 2015·Bioconjugate Chemistry·Dafni MoatsouRachel K O'Reilly
Dec 21, 2010·Journal of Biomedical Materials Research. Part a·William Monty ReichertDavid Tirrell
Nov 18, 2014·Nature Materials·Isidro CoboSébastien Perrier
Jan 24, 2008·Macromolecular Bioscience·Xiao-Wen ShiGregory F Payne
May 23, 2012·International Journal of Stomatology & Occlusion Medicine·Bogusława Orzechowska-WylęgałaBarbara Kalita
May 8, 2004·The Journal of Biological Chemistry·Henri R NordlundMarkku S Kulomaa
Oct 14, 2004·Journal of the American Chemical Society·Sivanand S PennadamCameron Alexander

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