Molecular adaptation of Drosophila melanogaster lysozymes to a digestive function.
A lysozyme (pI 5.5) was purified to homogeneity from heated acid extracts of Drosophila melanogaster larvae, using gel filtration in a Superose column and ion-exchange chromatography in a Mono Q column. The final yield was 67%. The purified lysozyme with Mr 13,700 (determined by SDS-polyacrylamide gel electrophoresis) decreases in activity and has its pH optimum displaced towards acidic values and Km increases as the ionic strength of the medium becomes higher. The lysozyme is resistant to a cathepsin D-like proteinase present in cyclorrhaphous Diptera and displays a chitinase activity which is 11-fold higher than that of chicken lysozyme. Microsequencing of an internal peptide of the purified lysozyme showed that this enzyme is the product of the previously sequenced Lys D gene. The results suggest that the product of the Lys P gene has pI 7.2, a pH optimum around 5 and is not a true digestive enzyme. The most remarkable sequence convergence of D. melanogaster lysozyme D and lysozymes from vertebrate foregut fermenters are serine 104 and a decrease in the number of basic amino acids, suggesting that these features are necessary for digestive function in an acid environment. Adaptive residues putatively conferring stability in an acid proteolytic environment differ between insects and vertebrates, probably because they depend on the overall three-dimensional structure of the lysozymes. A maximum likelihood phylogeny and inferences from insect lysozyme sequences showed that the recruitment of lysozymes as digestive enzymes is an ancestral condition of the flies (Diptera: Cyclorrhapha).
Departamento de Bioquímica, Universidade de São Paulo, Brasil.
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D000222:Adaptation, Physiological; D000595:Amino Acid Sequence; D000818:Animals; D004063:Digestion / Q000502:physiology*; D004064:Digestive System / Q000201:enzymology / Q000382:microbiology; D004331:Drosophila melanogaster / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D017344:Genes, Insect; D006801:Humans; D007526:Isoelectric Point; D007700:Kinetics; D007814:Larva / Q000201:enzymology; D008969:Molecular Sequence Data; D009113:Muramidase / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D010802:Phylogeny; D017386:Sequence Homology, Amino Acid
D009113:Muramidase