Conformational isomerism can limit antibody catalysis.

The Journal of Biological Chemistry
Erik W DeblerIan A Wilson

Abstract

Ligand binding to enzymes and antibodies is often accompanied by protein conformational changes. Although such structural adjustments may be conducive to enzyme catalysis, much less is known about their effect on reactions promoted by engineered catalytic antibodies. Crystallographic and pre-steady state kinetic analyses of antibody 34E4, which efficiently promotes the conversion of benzisoxazoles to salicylonitriles, show that the resting catalyst adopts two interconverting active-site conformations, only one of which is competent to bind substrate. In the predominant isomer, the indole side chain of Trp(L91) occupies the binding site and blocks ligand access. Slow conformational isomerization of this residue, on the same time scale as catalytic turnover, creates a deep and narrow binding site that can accommodate substrate and promote proton transfer using Glu(H50) as a carboxylate base. Although 34E4 is among the best catalysts for the deprotonation of benzisoxazoles, its efficiency appears to be significantly limited by this conformational plasticity of its active site. Future efforts to improve this antibody might profitably focus on stabilizing the active conformation of the catalyst. Analogous strategies may also be rele...Continue Reading

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Citations

Oct 23, 2009·Proceedings of the National Academy of Sciences of the United States of America·Erik W DeblerIan A Wilson
Jun 12, 2012·Proceedings of the National Academy of Sciences of the United States of America·Olga KhersonskyDan S Tawfik
Jul 29, 2015·Biochemistry·Arren Bar-EvenDan S Tawfik
Dec 29, 2009·Journal of Molecular Biology·Olga KhersonskyDan S Tawfik
Feb 10, 2017·The Journal of Physical Chemistry. B·Beytullah OzgurOzlem Keskin
Apr 20, 2021·Frontiers in Molecular Biosciences·Mohammed M Al QaraghuliPaul A Mulheran

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