Histidine phosphorylation in metalloprotein binding sites.

Post-translational modifications (PTMs) are invaluable regulatory tools for the control of catalytic functionality, protein-protein interactions, and signaling pathways. Historically, the study of phosphorylation as a PTM has been focused on serine, threonine, and tyrosine residues. In contrast, the significance of mammalian histidine phosphorylation remains largely unexplored. This gap in knowledge regarding the molecular basis for histidine phosphorylation as a regulatory agent exists in part because of the relative instability of phosphorylated histidine as compared with phosphorylated serine, threonine and tyrosine. However, the unique metal binding abilities of histidine make it one of the most common metal coordinating ligands in nature, and it is interesting to consider how phosphorylation would change the metal coordinating ability of histidine, and consequently, the properties of the phosphorylated metalloprotein. In this review, we examine eleven metalloproteins that have been shown to undergo reversible histidine phosphorylation at or near their metal binding sites. These proteins are described with respect to their biological activity and structure, with a particular emphasis on how phosphohistidine may tune the primary coordination sphere and protein conformation. Furthermore, several common methods, challenges, and limitations of studying sensitive, high affinity metalloproteins are discussed.

Thermodynamic Analysis of Metal-Ligand Cooperativity of PNP Ru Complexes: Implications for CO2 Hydrogenation to Methanol and Catalyst Inhibition.

The hydrogenation of CO2 in the presence of amines to formate, formamides, and methanol (MeOH) is a promising approach to streamlining carbon capture and recycling. To achieve this, understanding how catalyst design impacts selectivity and performance is critical. Herein we describe a thorough thermochemical analysis of the (de)hydrogenation catalyst, (PNP)Ru-Cl (PNP = 2,6-bis(di-tert-butylphosphinomethyl)pyridine; Ru = Ru(CO)(H)) and correlate our findings to catalyst performance. Although this catalyst is known to hydrogenate CO2 to formate with a mild base, we show that MeOH is produced when using a strong base. Consistent with pKa measurements, the requirement for a strong base suggests that the deprotonation of a six-coordinate Ru species is integral to the catalytic cycle that produces MeOH. Our studies also indicate that the concentration of MeOH produced is independent of catalyst concentration, consistent with a deactivation pathway that is dependent on methanol concentration, not equivalency. Our temperature-dependent equilibrium studies of the dearomatized congener, (*PNP)Ru, with various H-X species (to give (PNP)Ru-X; X = H, OH, OMe, OCHO, OC(O)NMe2) reveal that formic acid equilibrium is approximately temperature-independent; relative to H2, it is more favored at elevated temperatures. We also measure the hydricity of (PNP)Ru-H in THF and show how subsequent coordination of the substrate can impact the apparent hydricity. The implications of this work are broadly applicable to hydrogenation and dehydrogenation catalysis and, in particular, to those that can undergo metal-ligand cooperativity (MLC) at the catalyst. These results serve to benchmark future studies by allowing comparisons to be made among catalysts and will positively impact rational catalyst design.