New sequence motifs in flavoproteins: evidence for common ancestry and tools to predict structure

We describe two new sequence motifs, present in several families of flavoproteins. The "GG motif" (RxGGRxxS/T) is found shortly after the betaalphabetadinucleotide-binding motif (DBM) in L-amino acid oxidases, achacin and aplysianin-A, monoamine oxidases, corticosteroid-binding proteins, and tryptophan 2-monooxygenases. Other disperse sequence similarities between these families suggest a common origin. A GG motif is also found in protoporphyrinogen oxidase and carotenoid desaturases and, reduced to the central GG doublet, in the THI4 protein, dTDP-4-dehydrorhamnose reductase, soluble fumarate reductase, steroid dehydrogenases, Rab GDP-dissociation inhibitor, and in most flavoproteins with two dinucleotide-binding domains (glutathione reductase, glutamate synthase, flavin-containing monooxygenase, trimethylamine dehydrogenase...). In the latter families, an "ATG motif" (oxhhhATG) is found in both the FAD- and NAD(P)H-binding domains, forming the fourth beta-strand of the Rossman fold and the connecting loop. On the basis of these and previously described motifs, we present a classification of dinucleotide-binding proteins that could also serve as an evolutionary scheme. Like the DBM, the ATG motif appears to predate the divergence of NAD(P)H- and FAD-binding proteins. We propose that flavoproteins have evolved from a well-differentiated NAD(P)H-binding protein. The bulk of the substrate-binding domain was formed by an insertion after the fourth beta-strand, either of a closely related NAD(P)H-binding domain or of a domain of completely different origin.