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If life without heme-Fe were at all possible, it would definitely be different. Indeed this complex and versatile iron-porphyrin macrocycle upon binding to different “globins” yields hemeproteins crucial to sustain a variety of vital functions, generally classified, for convenience, in a limited number of functional families. Over-and-above the array of functions briefly outlined below, the spectacular progress in molecular genetics seen over the last 30 years led to the discovery of many hitherto unknown novel hemeproteins in prokaryotes and eukaryotes. Here, we highlight a few basic aspects of the chemistry of the hemeprotein universe, in particular those that are relevant to the control of heme-Fe reactivity and specialization, as sculpted by a variety of interactions with the protein moiety.

Chlorin e6 (Ce6) and its derivatives are among the most important photosensitizers in photodynamic therapy. Due to their intense fluorescence, chlorins may also be used for diagnostics. However, low solubility in water and high tendency to aggregation restrict their medical use. Here we demonstrate that apo-myoglobin, by reinserting Ce6 in its heme binding pocket, can be used to monomolecularly disperse it. The reconstructed myoglobin-Ce6 adduct presents noticeable changes in the photophysical properties of the chromophore. A red-shift, in particular in the transparency window, can be observed in the absorption and in the emission spectra of the adduct compared to the spectra of the free chlorin in PBS. The adduct presents a higher quantum yield and an increased excited-state lifetime with respect to the free Ce6. The binding of Ce6 to apo-myoglobin determines a decrease of the ¹O₂ generation but a three-fold increase of peroxides production, determining globally an increase in the performance of Ce6 as a photosensitizer and imaging agent.

Nitroorganics present a general concern for a safe environment due to their health hazards. However, some nitroorganics such as metronidazole (Mtz) and chloramphenicol (CAM) also possess medicinal value. Mtz and CAM can undergo reductive bioactivation presumably via their nitroso derivatives. We show, using UV-vis spectroscopy, that sperm whale myoglobin (swMb) and its distal pocket mutants retaining H-bonding capacity react with Mtz in the presence of dithionite to generate products with spectra suggestive of the Fe-bound nitroso (Fe–RNO; ${\lambda }_{max}$$\sim$420 nm) forms. We have crystallized and solved the X-ray crystal structure of an H64Q swMb-acetamide compound to 1.76 Å resolution; the formation of this compound results from the serendipitous crystallographic trapping, by the heme center, of acetamide from the reductive decomposition of Mtz. Only one of the swMb proteins, namely H64Q swMb with a relatively flexible Gln64 residue, reacted with CAM presumably due to the bulky nature of CAM that generally may restrict its access to the heme site.

Sulfoxylate SO₂H⁻(SO₂²⁻), a strong reducing agent readily produced by hydrolysis of thiourea dioxide, reacts with ferric myoglobin (Mb) to reversibly produce Fe(II)-Mb, starting from either aerobic or anaerobic conditions. Exposure of Fe(II)-Mb to excess sulfoxylate further produces Fe(II)-CO-Mb. Fe(II)-Mb can be regenerated by reoxidation with ferricyanide at this stage; hemin, rubredoxin and cytochrome c show a similar reactivity towards sulfoxylate. The source of CO is not the protein moiety, nor is it the heme or the thiourea dioxide – but rather CO₂, via its reaction with sulfoxylate when the latter is used in large excess. These findings provide a convenient single-step route to carbon monoxide heme adducts, without the need to manipulate toxic CO gas.

Protoheme IX is a typical iron porphyrin cofactor, showing a variety of reactivities in many hemoproteins under the reaction environments provided by protein matrices. Chemical modification of the protoheme cofactor is expected to be a versatile strategy to design hemoproteins possessing unique functions. This review focuses on the conversion of a hemoprotein, mainly myoglobin (an oxygen-storage hemoprotein), into a protein having different functions from the original ones by replacement of the protoheme cofactor with synthetic cofactors. The myoglobin having anionic patches pended to the heme propionates effectively binds electron-accepting proteins or small cationic organic molecules on the protein surface, resulting in enhanced efficiency of the photoinduced electron transfers from the myoglobin to these electron acceptors. Furthermore, the peroxidase and peroxygenase activities are also enhanced due to the facile substrate accesses. The attachment of the chemically active moiety such as flavin at the heme terminal is also important to give P450-like function to the native myoglobin. The employment of a structural isomer of porphyrin as an artificial cofactor gives rise to remarkably high dioxygen affinity and peroxidase activity in myoglobin, and allows us to easily detect high-valent species of the porphyrin isomer in HRP. These examples provide a clear insight into hemoprotein modifications based on synthetic chemistry as well as genetic amino acid mutations.

Continuous-wave linear-absorption spectroscopy based on THz radiation generated by solid-state photomixers has been applied to the investigation of the dynamics of biomolecules in polyethylene matrices and to line shape studies of HF for diagnostics of semiconductor etching plasmas. The THz spectra of biotin and myoglobin have been obtained using a variable-temperature, cryogenic sampling system. The spectrum of biotin displays a small number of discrete absorptions over the temperature range from 4.2 K to room temperature while the spectrum of myoglobin has no obvious resonance structure at the >10% fractional absorption level. Spectral predictions from the lowest energy ab initio conformations of biotin are in poor agreement with experiment, suggesting the need to include condensed-phase environmental interactions for qualitative predictions of the THz spectrum. Vibrational anharmonicity is used to model the line shapes that result from drastic changes in vibrational sequence level populations of biotin over this temperature range. Anharmonicity factors (χ_eω_e/ω_e) at the levels of 0.1 % to 0.8 % are obtained from non-linear least squares fits of the observed resonances and illustrate their important for refining model predictions. Application of the photomixer system to line shape studies in etching plasmas has been used to study the formation efficiency and translational temperature of HF at 1.2 THz under different operating conditions. These results will aid in understanding the chemistry of industry-standard fluorocarbon and oxygenated fluorocarbon etching plasmas.