التفاصيل البيبلوغرافية
العنوان: |
How to Build a Metalloenzyme: Lessons from a Protein-Based Model of Acetyl Coenzyme A Synthase. |
المؤلفون: |
Shafaat HS, Manesis AC; Departments of Molecular Biosciences and Chemistry, Northwestern University, Evanston, Illinois 60208, United States., Yerbulekova A |
المصدر: |
Accounts of chemical research [Acc Chem Res] 2023 May 02; Vol. 56 (9), pp. 984-993. Date of Electronic Publication: 2023 Apr 12. |
نوع المنشور: |
Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, Non-U.S. Gov't |
اللغة: |
English |
بيانات الدورية: |
Publisher: American Chemical Society Country of Publication: United States NLM ID: 0157313 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1520-4898 (Electronic) Linking ISSN: 00014842 NLM ISO Abbreviation: Acc Chem Res Subsets: MEDLINE |
أسماء مطبوعة: |
Original Publication: Washington, American Chemical Society. |
مواضيع طبية MeSH: |
Azurin*/metabolism , Metalloproteins*, Acetyl Coenzyme A/chemistry ; Acetyl Coenzyme A/metabolism ; Nickel/chemistry ; Carbon Monoxide/metabolism ; Electron Spin Resonance Spectroscopy |
مستخلص: |
"What I cannot create, I do not understand"─Richard Feynman. This sentiment motivates the entire field of artificial metalloenzymes. Naturally occurring enzymes catalyze reactions with efficiencies, rates, and selectivity that generally cannot be achieved in synthetic systems. Many of these processes represent vital building blocks for a sustainable society, including CO 2 conversion, nitrogen fixation, water oxidation, and liquid fuel synthesis. Our inability as chemists to fully reproduce the functionality of naturally occurring enzymes implicates yet-unknown contributors to reactivity. To identify these properties, it is necessary to consider all of the components of naturally occurring metalloenzymes, from the active site metal(s) to large-scale dynamics. In this Account, we describe the holistic development of a metalloprotein-based model that functionally reproduces the acetyl coenzyme A synthase (ACS) enzyme.ACS catalyzes the synthesis of a thioester, acetyl coenzyme A, from gaseous carbon monoxide, a methyl group donated by a cobalt corrinoid protein, and coenzyme A. The active site of ACS contains a bimetallic nickel site coupled to a [4Fe-4S] cluster. This reaction mimics Monsanto's acetic acid synthesis and represents an ancient process for incorporating inorganic carbon into cellular biomass through the primordial Wood-Ljungdahl metabolic pathway. From a sustainability standpoint, the reversible conversion of C 1 substrates into an acetyl group and selective downstream transfer to a thiolate nucleophile offer opportunities to expand this reactivity to the anthropogenic synthesis of liquid fuels. However, substantial gaps in our understanding of the ACS catalytic mechanism coupled with the enzyme's oxygen sensitivity and general instability have limited these applications. It is our hope that development of an artificial metalloenzyme that carries out ACS-like reactions will advance our mechanistic understanding and enable synthesis of robust compounds with the capacity for similar reactivity.To construct this model, we first focused on the catalytic proximal nickel (Ni P ) site, which has a single metal center bound by three bridging cysteine residues in a "Y"-shaped arrangement. With an initial emphasis on reproducing the general structure of a low-coordinate metal binding site, the type I cupredoxin, azurin, was selected as the protein scaffold, and a nickel center was incorporated into the mononuclear site. Using numerous spectroscopic and computational techniques, including electron paramagnetic resonance (EPR) spectroscopy, nickel-substituted azurin was shown to have similar electronic and geometric structures to the Ni P center in ACS. A substrate access channel was installed, and both carbon monoxide and a methyl group were shown to bind individually to the reduced Ni I center. The elusive EPR-active S = 1/2 Ni-CH 3 species, which has never been detected in native ACS, was observed in the azurin-based model, establishing the capacity of a biological Ni I species to support two-electron organometallic reactions. Pulsed EPR studies on the S = 1/2 Ni-CH 3 species in azurin suggested a noncanonical electronic structure with an inverted ligand field, which was proposed to prevent irreversible site degradation. This model azurin protein was ultimately shown to perform carbon-carbon and carbon-sulfur bond formation using sequential, ordered substrate addition for selective, stoichiometric thioester synthesis. X-ray spectroscopic methods were used to provide characterization of the remaining catalytic intermediates, resolving some debate over key mechanistic details.The overall approach and strategies that we employed for the successful construction of a functional protein-based model of ACS are described in this Account. We anticipate that these principles can be adapted across diverse metalloenzyme classes, providing essential mechanistic details and guiding the development of next-generation, functional artificial metalloenzymes. |
المشرفين على المادة: |
12284-43-4 (Azurin) 72-89-9 (Acetyl Coenzyme A) 7OV03QG267 (Nickel) 7U1EE4V452 (Carbon Monoxide) 0 (Metalloproteins) |
تواريخ الأحداث: |
Date Created: 20230412 Date Completed: 20230503 Latest Revision: 20230517 |
رمز التحديث: |
20231215 |
DOI: |
10.1021/acs.accounts.2c00824 |
PMID: |
37042748 |
قاعدة البيانات: |
MEDLINE |