glutamyl-glutamic-acid and aspartylglutamate

Phosphopantetheinyl transferases (PPTases), which play an essential role in both primary and secondary metabolism, are magnesium binding enzymes. In this study, we characterized the magnesium binding residues of all known group II PPTases by biochemical and evolutionary analysis. Our results suggested that group II PPTases could be classified into two subgroups, two-magnesium-binding-residue-PPTases containing the triad Asp-Xxx-Glu and three-magnesium-binding-residue-PPTases containing the triad Asp-Glu-Glu. Mutations of two three-magnesium-binding-residue-PPTases and one two-magnesium-binding-residue-PPTase indicate that the first and the third residues in the triads are essential to activities; the second residues in the triads are non-essential. Although variations of the second residues in the triad Asp-Xxx-Glu exist throughout the whole phylogenetic tree, the second residues are conserved in animals, plants, algae, and most prokaryotes, respectively. Evolutionary analysis suggests that: the animal group II PPTases may originate from one common ancestor; the plant two-magnesium-binding-residue-PPTases may originate from one common ancestor; the plant three-magnesium-binding-residue-PPTases may derive from horizontal gene transfer from prokaryotes.

Topics: Amino Acid Sequence; Bacterial Proteins; Binding Sites; Biological Evolution; Dipeptides; Magnesium; Molecular Sequence Data; Phylogeny; Sequence Alignment; Transferases (Other Substituted Phosphate Groups)

Aspartate binds calcium(II) better than glutamate with Ka = 7.0 ± 0.9 L mol⁻¹ for Asp and Ka = 3.0 ± 0.8 L mol⁻¹ for Glu, respectively, as determined using calcium-selective electrodes for aqueous solutions of ionic strength 0.20 at 25 °C at pH of relevance for milk products. For the mixed peptides, the affinity seems additive with Ka = 27 ± 3 L mol⁻¹ for Asp-Glu and 22.7 ± 0.1 for Glu-Asp as compared to the expected 21 L mol⁻¹. In contrast, for Asp-Asp, the affinity is less than additive with Ka = 23 ± 5 L mol⁻¹ as compared to the expected 49 L mol⁻¹, whereas for Glu-Glu, the affinity is more than additive with Ka = 26 ± 4 L mol⁻¹ as compared to the expected 9.0 L mol⁻¹, indicating specific structural effects for Glu-Glu. Ionic strength effects, 1.0 versus 0.20 studied, are similar for Asp and Glu with decreasing affinity for higher ionic strength, whereas the dipeptides with Glu as C-terminus are more sensitive to increasing ionic strength than with Asp as C-terminus. Despite little affinity of calcium to serine with Ka = 0.9 ± 0.2 L mol⁻¹, Glu has increasing affinity for calcium in the serine dipeptide Ser-Glu with Ka = 10 ± 3 L mol⁻¹, which becomes comparable to phosphorylated serine with Ka = 22 ± 5 L mol⁻¹.

Topics: Beverages; Calcium; Calcium, Dietary; Chelating Agents; Dipeptides; Food Additives; Food, Fortified; Humans; Hydrogen-Ion Concentration; Kinetics; Nutritive Value