Making or Breaking Metal-Dependent Catalytic Activity: The Role of Stammers in Designed Three-Stranded Coiled Coils.

Autor: Pinter TBJ; Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA., Manickas EC; Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA., Tolbert AE; Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA., Koebke KJ; Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA., Deb A; Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA., Penner-Hahn JE; Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA., Pecoraro VL; Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA.
Jazyk: angličtina
Zdroj: Angewandte Chemie (International ed. in English) [Angew Chem Int Ed Engl] 2020 Nov 09; Vol. 59 (46), pp. 20445-20449. Date of Electronic Publication: 2020 Sep 02.
DOI: 10.1002/anie.202008356
Abstrakt: While many life-critical reactions would be infeasibly slow without metal cofactors, a detailed understanding of how protein structure can influence catalytic activity remains elusive. Using de novo designed three-stranded coiled coils (TRI and Grand peptides formed using a heptad repeat approach), we examine how the insertion of a three residue discontinuity, known as a stammer insert, directly adjacent to a (His) 3 metal binding site alters catalytic activity. The stammer, which locally alters the twist of the helix, significantly increases copper-catalyzed nitrite reductase activity (CuNiR). In contrast, the well-established zinc-catalyzed carbonic anhydrase activity (p-nitrophenyl acetate, pNPA) is effectively ablated. This study illustrates how the perturbation of the protein sequence using non-coordinating and non-acid base residues in the helical core can perturb metalloenzyme activity through the simple expedient of modifying the helical pitch adjacent to the catalytic center.
(© 2020 Wiley-VCH GmbH.)
Databáze: MEDLINE
Externí odkaz: