Roles of hierarchical and metabolic regulation in the allometric scaling of metabolism in Panamanian orchid bees

Abstract

Assessment of the relative importance of variation in enzyme concentration [E] and metabolic regulation in accounting for interspecific variation in metabolic rates is an unrealized area of research. Towards this end, we used metabolic flux rates during hovering and enzymatic flux capacities (V max values, equal to [E]Xkcat, where kcat is catalytic efficiency) in flight muscles measured in vitro from 14 orchid bee species ranging in body mass from 47 to 1065 mg. Previous studies revealed that, across orchid bee species, wingbeat frequencies and metabolic rates decline in parallel with increasing body mass. Vmax values at some enzymatic steps in pathways of energy metabolism decline with increasing mass while, at most other steps, Vmax values are mass-independent. We quantified the relative importance of 'hierarchical regulation' (alteration in Vmax, indicative of alteration in [E]) and 'metabolic regulation' (resulting from variation in substrate, product or modulator concentrations) in accounting for interspecific variation in flux across species. In addition, we applied the method of phylogenetically independent contrasts to remove the potentially confounding effects of phylogenetic relationships among species. In the evolution of orchid bees, hierarchical regulation completely accounts for allometric variation in flux rates at the hexokinase step while, at other reactions, variation in flux is completely accounted for by metabolic regulation. The predominant role played by metabolic regulation is examined at the phosphoglucoisomerase step using the Haldane relationship. We find that extremely small variation in the concentration ratio of [product]/[substrate] is enough to cause the observed interspecific variation in net flux at this reaction in glycolysis.