tryptophan-tryptophylquinone and quinone

MauG catalyzes posttranslational modifications of a methylamine dehydrogenase precursor (preMADH) to complete the biosynthesis of its protein-derived tryptophan tryptophylquinone (TTQ) cofactor. Trp199 is present at the site of interaction between MauG and preMADH and is critical to this process as it mediates hole hopping during the inter-protein electron transfer that is required for catalysis. Trp199 was converted to Glu and the structure and reactivity of the W199E/preMADH complex were characterized. The results reveal that the nature of residue 199 is also important for productive complex formation between preMADH and MauG.

Topics: Bacterial Proteins; Benzoquinones; Electron Transport; Enzyme Precursors; Indolequinones; Iron; Models, Molecular; Mutagenesis, Site-Directed; Mutation; Oxidoreductases Acting on CH-NH Group Donors; Paracoccus denitrificans; Protein Binding; Protein Conformation; Tryptophan

Five mutants of Methylobacterium extorquens and four mutants of Paracoccus denitrificans that have a general defect in c-type cytochrome synthesis also failed to assemble an active methylamine dehydrogenase. In all cases methanol dehydrogenase, another periplasmic enzyme, was fully active. All nine mutant strains accumulated both the heavy and light subunits of methylamine dehydrogenase to essentially wild-type levels. In all nine mutants, the heavy-subunit and light-subunit polypeptides were proteolytically processed, suggesting that translocation to the periplasm had occurred; in the case of the P. denitrificans mutants, a periplasmic location for the heavy and light subunits was confirmed experimentally. While specific quinone staining of the methylamine dehydrogenase light subunit in wild-type M. extorquens and P. denitrificans strains could readily be demonstrated, the light subunit polypeptides accumulated by the mutants did not quinone stain, indicating that the methylamine dehydrogenase prosthetic group, tryptophan tryptophylquinone, is not assembled in the absence of functional c-type cytochromes.

Topics: Benzoquinones; Choline; Cytochrome c Group; Gram-Negative Aerobic Bacteria; Indolequinones; Methylamines; Mutation; Oxidoreductases Acting on CH-NH Group Donors; Paracoccus denitrificans; Peptides; Quinones; Tryptophan