Tyrosine Latching of a Regulatory Gate Affords Allosteric Control of Aromatic Amino Acid Biosynthesis*

The first step of the shikimate pathway for aromatic amino acid biosynthesis is catalyzed by 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase (DAH7PS). Thermotoga maritima DAH7PS (TmaDAH7PS) is tetrameric, with monomer units comprised of a core catalytic (β/α)8 barrel and an N-terminal domain. This enzyme is inhibited strongly by tyrosine and to a lesser extent by the presence of phenylalanine. A truncated mutant of TmaDAH7PS lacking the N-terminal domain was catalytically more active and completely insensitive to tyrosine and phenylalanine, consistent with a role for this domain in allosteric inhibition. The structure of this protein was determined to 2.0 Å. In contrast to the wild-type enzyme, this enzyme is dimeric. Wild-type TmaDAH7PS was co-crystallized with tyrosine, and the structure of this complex was determined to a resolution of 2.35 Å. Tyrosine was found to bind at the interface between two regulatory N-terminal domains, formed from diagonally located monomers of the tetramer, revealing a major reorganization of the regulatory domain with respect to the barrel relative to unliganded enzyme. This significant conformational rearrangement observed in the crystal structures was also clearly evident from small angle X-ray scattering measurements recorded in the presence and absence of tyrosine. The closed conformation adopted by the protein on tyrosine binding impedes substrate entry into the neighboring barrel, revealing an unusual tyrosine-controlled gating mechanism for allosteric control of this enzyme

Was Phanerozoic reef history controlled by the distribution of non-enzymatically secreted reef carbonates (microbial carbonate and biologically induced cement)?

Throughout most of the Phanerozoic, reef rigidity resulted as much, or more, from early lithification by microbial carbonates and biologically induced cements (non-enzymatic carbonates) than from biological encrustation of, or by, large, enzymatically secreted metazoan skeletons. Reef framework is divided into four categories: (1) skeletal metazoan; (2) non-skeletal microbialite (stromatolite and thrombolite); (3) calcimicrobe; and (4) biocementstone, in which small or delicate organisms serve as scaffolds for rigid cement crusts. The last three categories are dominated by non-enzymatic carbonates. Skeletal framework and non-skeletal microbialite framework were the most abundant framework types through the Phanerozoic. The composition and abundance of skeletal framework was controlled largely by mass extinction events, but most reefs consisted of both microbialite and skeletal organisms in a mutually beneficial relationship. Microbialite framework was abundant throughout the Palaeozoic and early Mesozoic, but declined after the Jurassic. Calcimicrobe framework was important during the Cambrian-Early Ordovician and Devonian and biocementstone framework was important from the late Mississippian to the Late Triassic. The Phanerozoic history of reefs does not correlate well with the stratigraphic distribution of large, skeletal 'reef builders', or with a variety of physicochemical parameters, including sea-level history, Wilson Cycle or global climate cycles. Because non-enzymatic carbonates result from induction by non-obligate calcifiers, and not enzymatic precipitation by obligate calcifiers, the distribution of these carbonates was controlled to a larger extent by temporal changes in physicochemical parameters affecting the saturation state of sea water with respect to carbonate minerals. Changes in pCO, Ca/Mg ratios, cation concentrations and temperature may have affected the abundance of non-enzymatic carbonates and, hence, reefs, independently from the effects of these same parameters and mass extinction events on skeletal reef biota. The decline in abundance of reefal microbialite and absence of calcimicrobe and biocementstone reef framework after the Jurassic may be a result of relatively low saturation states of sea water owing to increased removal and sequestration of finite marine carbonate resources by calcareous plankton since the Jurassic. Reef history is difficult to correlate with temporal changes in specific global parameters because these parameters affect skeletal biota and biologically induced carbonate precipitation independently. Hence, reef history was regulated not just by skeletal reef biota, but by parameters governing non-enzymatic carbonates