Artificial Metalloproteins: At the Interface between Biology and Chemistry.

Artificial metalloproteins (ArMs) have recently gained significant interest due to their potential to address issues in a broad scope of applications, including biocatalysis, biotechnology, protein assembly, and model chemistry. ArMs are assembled by the incorporation of a non-native metallocofactor into a protein scaffold. This can be achieved by a number of methods that apply tools of chemical biology, computational de novo design, and synthetic chemistry. In this Perspective, we highlight select systems in the hope of demonstrating the breadth of ArM design strategies and applications and emphasize how these systems address problems that are otherwise difficult to do so with strictly biochemical or synthetic approaches.

Scaffold-based [Fe]-hydrogenase model: H2 activation initiates Fe(0)-hydride extrusion and non-biomimetic hydride transfer.

We report the synthesis and reactivity of a model of [Fe]-hydrogenase derived from an anthracene-based scaffold that includes the endogenous, organometallic acyl(methylene) donor. In comparison to other non-scaffolded acyl-containing complexes, the complex described herein retains molecularly well-defined chemistry upon addition of multiple equivalents of exogenous base. Clean deprotonation of the acyl(methylene) C-H bond with a phenolate base results in the formation of a dimeric motif that contains a new Fe-C(methine) bond resulting from coordination of the deprotonated methylene unit to an adjacent iron center. This effective second carbanion in the ligand framework was demonstrated to drive heterolytic H2 activation across the Fe(ii) center. However, this process results in reductive elimination and liberation of the ligand to extrude a lower-valent Fe-carbonyl complex. Through a series of isotopic labelling experiments, structural characterization (XRD, XAS), and spectroscopic characterization (IR, NMR, EXAFS), a mechanistic pathway is presented for H2/hydride-induced loss of the organometallic acyl unit (i.e. pyCH2-C[double bond, length as m-dash]O → pyCH3+C[triple bond, length as m-dash]O). The known reduced hydride species [HFe(CO)4]- and [HFe3(CO)11]- have been observed as products by 1H/2H NMR and IR spectroscopies, as well as independent syntheses of PNP[HFe(CO)4]. The former species (i.e. [HFe(CO)4]-) is deduced to be the actual hydride transfer agent in the hydride transfer reaction (nominally catalyzed by the title compound) to a biomimetic substrate ([TolIm](BArF) = fluorinated imidazolium as hydride acceptor). This work provides mechanistic insight into the reasons for lack of functional biomimetic behavior (hydride transfer) in acyl(methylene)pyridine based mimics of [Fe]-hydrogenase.

Scaffold-Based Functional Models of [Fe]-Hydrogenase (Hmd): Building the Bridge between Biological Structure and Molecular Function.

The well-known dinuclear [FeFe] and [NiFe] hydrogenase enzymes are redox-based proton reduction and H2 oxidation catalysts. In comparison, the structural and functional aspects of the mononuclear nonredox hydrogenase, known as [Fe]-hydrogenase or Hmd, have been less explored because of the relatively recent crystallographic elucidation of the enzyme active site. Additionally, the synthetic challenges posed by the highly substituted and asymmetric coordination environment of the iron guanylylpyridinol (FeGP) cofactor have hampered functional biomimetic modeling studies to a large extent. The active site contains an octahedral low-spin Fe(II) center with the following coordination motifs: a bidentate acyl-pyridone moiety (C,N) and cysteinyl-S in a facial arrangement; two cis carbonyl ligands; and a H2O/H2 binding site. In [Fe]-hydrogenase, heterolytic H2 activation putatively by the pendant pyridone/pyridonate-O base serving as a proton acceptor. Following H2 cleavage, an intermediate Fe-H species is thought to stereoselectively transfer a hydride to the substrate methenyl-H4MPT+, thus forming methylene-H4MPT. In the past decade, chemists, inspired by the elegant organometallic chemistry inherent to the FeGP cofactor, have synthesized a number of faithful structural models. However, functional systems are still relatively limited and often rely on abiological ligands or metal centers that obfuscate a direct correlation to nature's design.Our group has developed a bioinspired suite of synthetic analogues of Hmd to better understand the effects of structure on the stability and functionality of the Hmd active site, with a special emphasis on using a scaffold-based ligand design. This systematic approach has contributed to a deeper understanding of the unique ligand array of [Fe]-hydrogenase in nature and has ultimately resulted in the first functional synthetic models without the aid of abiological ligands. This Account reviews the reactivity of the functional anthracene-scaffolded synthetic models developed by our group in the context of current mechanistic understanding drawn from both protein crystallography and computational studies. Furthermore, we introduce a novel thermodynamic framework to place the reactivity of our model systems in context and provide an outlook on the future study of [Fe]-hydrogenase synthetic models through both a structural and functional lens.

Bioinspired CNP Iron(II) Pincers Relevant to [Fe]-Hydrogenase (Hmd): Effect of Dicarbonyl versus Monocarbonyl Motifs in H2 Activation and Transfer Hydrogenation.

A set of bioinspired carbamoyl CNP pincer complexes are reported that are relevant to [Fe]-hydrogenase (Hmd). The dicarbonyl species [(CNHNNHPR2)Fe(CO)2I] [R = Ph, 1; R = iPr, 2] undergoes ligand deprotonation, resulting in the dearomatized complexes of formulas [(CNHNN=PR2)Fe(CO)2] (5 and 6). The crystal structure and 1H{31P} NMR spectroscopy of the iodide-bound dearomatized species [Na(18-crown-6)][(CNHNN=PPh2)Fe(CO)2I] (7) showed that the deprotonated moiety was the phosphoramine N(H) linkage. Separately, the monocarbonyl complexes [(CNHNNHPR2)Fe(CO)(MeCN)2](BF4) (8 and 9) synthesized, as well as deprotonated and dearomatized in similar fashion. Reactivity studies revealed that the parent dicarbonyl complexes require more forceful conditions for H2 activation, compared with the monocarbonyl complexes. The ligand backbone was not found to participate in H2 activation and H2 → hydride transfer to an organic substrate was not observed in either case. Density functional theory calculations revealed that the higher reactivity of the monocarbonyl complex in H2 splitting could be attributed to its higher affinity for H2. This behavior is attributed to two key points related to the requisite dπ(Fe) → σ*(H2) back-bonding interaction in a conventional M-H2 Kubas interaction: (i) generally, the weaker π donor capacity of the dicarbonyls, and (ii) specifically, the detrimental effect of a strongly π acidic CO ligand (versus weakly π acidic MeCN ligand) trans to the H2 activation site. The higher reactivity of the monocarbonyl complex is also evidenced by the catalytic transfer hydrogenation by monocarbonyl 8, whereas dicarbonyl 1 was ineffective. Overall, the results suggest that Nature uses the dicarbonyl motif in [Fe]-hydrogenase to diminish the interaction between the Fe center and dihydrogen, thereby preventing premature H2 activation prior to substrate (H4MPT+) binding and any resulting nonspecific hydride transfer reactivity.

Functional Hydride Transfer by a Thiolate-Containing Model of Mono-Iron Hydrogenase featuring an Anthracene Scaffold.

We report the synthesis, X-ray structure and functional biomimetic activity of a model complex of mono-iron hydrogenase (Hmd). To achieve the desired biomimetic fac-CNS(thiolate) ligation motif, an anthracene framework is used to provide the requisite donors in a single chelate. A bulky aryl thiolate (ortho dimethylphenyl) is included to achieve mononuclearity. In addition to exhibiting structural (X-ray) and spectroscopic (NMR, IR) similarity to the enzyme, the complex is competent for H2 activation (heterolysis) and hydride transfer to a model substrate-mimicking the functional behavior of the enzyme in a biomimetic CNS coordination sphere for the first time.