Nitrite reductase activity within an antiparallel de novo scaffold.

Autor: Koebke KJ; Department of Chemistry, University of Michigan, Ann Arbor, MI, USA., Tebo AG; Program in Chemical Biology, University of Michigan, Ann Arbor, MI, USA.; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA., Manickas EC; Department of Chemistry, University of Michigan, Ann Arbor, MI, USA., Deb A; Department of Chemistry, University of Michigan, Ann Arbor, MI, USA., Penner-Hahn JE; Department of Chemistry, University of Michigan, Ann Arbor, MI, USA.; Department of Biophysics, University of Michigan, Ann Arbor, MI, USA., Pecoraro VL; Department of Chemistry, University of Michigan, Ann Arbor, MI, USA. vlpec@umich.edu.
Jazyk: angličtina
Zdroj: Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry [J Biol Inorg Chem] 2021 Oct; Vol. 26 (7), pp. 855-862. Date of Electronic Publication: 2021 Sep 06.
DOI: 10.1007/s00775-021-01889-1
Abstrakt: Copper nitrite reductase (CuNiR) is a copper enzyme that converts nitrite to nitric oxide and is an important part of the global nitrogen cycle in bacteria. The relatively simple CuHis 3 binding site of the CuNiR active site has made it an enticing target for small molecule modeling and de novo protein design studies. We have previously reported symmetric CuNiR models within parallel three stranded coiled coil systems, with activities that span a range of three orders of magnitude. In this report, we investigate the same CuHis 3 binding site within an antiparallel three helical bundle scaffold, which allows the design of asymmetric constructs. We determine that a simple CuHis 3 binding site can be designed within this scaffold with enhanced activity relative to the comparable construct in parallel coiled coils. Incorporating more complex designs or repositioning this binding site can decrease this activity as much as 15 times. Comparing these constructs, we reaffirm a previous result in which a blue shift in the 1s to 4p transition energy determined by Cu(I) X-ray absorption spectroscopy is correlated with an enhanced activity within imidazole-based constructs. With this step and recent successful electron transfer site designs within this scaffold, we are one step closer to a fully functional de novo designed nitrite reductase.
(© 2021. Society for Biological Inorganic Chemistry (SBIC).)
Databáze: MEDLINE
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