Investigations into the mechanism of orotidine 5'-monophosphate decarboxylase: Sources of substrate destabilization and transition state stabilization

Orotidine 5′-monophosphate decarboxylase (OMPDC) achieves a rarely paralleled rate acceleration, yet the catalytic basis prompting this enhancement have yet to be fully elucidated. To accomplish decarboxylation, OMPDC must overcome the high energy barrier due to the localized anionic charge of the intermediate. Mechanistic studies employing enzyme mutagenesis and product or intermediate analogues were used to investigate possible transition state stabilization by a carbene resonance structure. Viability of the carbene structure depends upon a key hydrogen bond between O4 of the substrate and the amide backbone of a conserved serine or threonine. Substitution of the conserved residue with Pro resulted in a kcat/KM of 1 M-1s-1; deletion of the FUMP O4 resulted in a product analogue that does not undergo H6 exchange or inhibit decarboxylation. Hence, indirect evidence reveals the O4-backbone interaction plays an important role for binding and catalysis. OMPDC likely has honed multiple mechanisms to attain its remarkable catalysis. The successful crystallizations of OMPDC a decade ago sparked hypotheses that structure and sequence conserved residues induced productive strain on the substrate-enzyme complex. Here, we demonstrate a new source of stress: a hydrophobic pocket adjacent to the OMP carboxylate that exhibits kinetic parameters characteristic of substrate destabilization. Substitution of these residues with hydrophilic side-chains, by providing hydrogen-bonding partners, decreased kcat by 10 to 10^4–fold. The same substitutions display very little change in the rate of product H6 exchange, supporting that this hydrophobic pocket affects the substrate-enzyme complex before the transition state. We also provide evidence that hydrophilic residues can insert water molecules into the pocket with detrimental effects to catalysis.

Consumerism in urban Russia in the 1880s and 90s: Exploring conceptions of time, space, and modernity through bicycle and watch advertisements

This paper discusses the urban consumer culture in Moscow and Petersburg during the 1880s and 1890s and uses the consumption of bicycles and watches as a lens through which to explore changing perceptions of time and space within the experience of modernity at the end of the nineteenth century. Specifically, I argue that the way in which consumers and merchants constructed a dialogue of meaning around particular objects; the way in which objects are consumed by a culture gives insight into the values, morals, and tenure of that culture. The paper preferences newspaper ads and photographs as the mouthpieces of merchants and consumers respectively as they constructed a dialogue in the language of consumerism, and explores the ways in which both parties sought to assign meaning to objects during the experience of modernity. I am particularly interested in the way consumers perform elements of cultural modernity in photographs and how these instances of performance relate to their negotiation of modernity. The paper takes as its focus large section of the urban Russian population, much of whom can traditionally be called “middle class” but whose diversity has led me to the adoption of the term “consumer community,” and whose makeup is described in detail. The paper contributes to the continuing scholarly discourse on the makeup of the middle class in Russia and the social boundaries of late tsarist society. It speaks to the the developing sensibilities and values of a generation struggling to define itself in a rapidly changing world, to the ways in which conceptualizations of public and private space, as well as feminine and masculine space were redefined, and to the developing visual culture of the Russian consumer society, largely predicated on the display of objects to signify socially desirable traits. Whereas other explorations of consumer culture and advertisements have portrayed the relationship between merchants and consumers as a one-sided monologue in which merchants convince consumers that certain objects have cultural value, I emphasis the dialogue between merchants and consumers, and their mutual negotiation of cultural meaning through objects.