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Making or Breaking Metal‐Dependent Catalytic Activity: The Role of Stammers in Designed Three‐Stranded Coiled Coils
Author(s) -
Pinter Tyler B. J.,
Manickas Elizabeth C.,
Tolbert Audrey E.,
Koebke Karl J.,
Deb Aniruddha,
PennerHahn James E.,
Pecoraro Vincent L.
Publication year - 2020
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.202008356
Subject(s) - chemistry , catalysis , metal , zinc , stereochemistry , cofactor , crystallography , combinatorial chemistry , biochemistry , enzyme , organic chemistry
Abstract 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.