title: Role of a single amino acid in the evolution of glycans of invertebrates and vertebrates.
abstract: Structures of glycoconjugate N-glycans and glycolipids of invertebrates show significant differences from those of vertebrates. These differences are due largely to the vertebrate beta1,4-galactosyltransferase-1 (beta4Gal-T1), which is found as a beta1,4-N-acetylgalactosaminyltransferase (beta4GalNAc-T1) in invertebrates. Mutation of Tyr285 to Ile or Leu in human beta4Gal-T1 converts the enzyme into an equally efficient beta4GalNAc-T1. A comparison of all the human beta4Gal-T1 ortholog enzymes shows that this Tyr285 residue in human beta4Gal-T1 is conserved either as Tyr or Phe in all vertebrate enzymes, while in all invertebrate enzymes it is conserved as an Ile or Leu. We find that mutation of the corresponding Ile residue to Tyr in Drosophila beta4GalNAc-T1 converts the enzyme to a beta4Gal-T1 by reducing its N-acetylgalactosaminyltransferase activity by nearly 1000-fold, while enhancing its galactosyltransferase activity by 80-fold. Furthermore, we find that, similar to the vertebrate/mammalian beta4Gal-T1 enzymes, the wild-type Drosophila beta4GalNAc-T1 enzyme binds to a mammary gland-specific protein, alpha-lactalbumin (alpha-LA). Thus, it would seem that, during the evolution of vertebrates from invertebrates over 500 million years ago, beta4Gal-T1 appeared as a result of the single amino acid substitution of Tyr or Phe for Leu or Ile in the invertebrate beta4GalNAc-T1. Subsequently, the pre-existing alpha-LA-binding site was utilized during mammalian evolution to synthesize lactose in the mammary gland during lactation.
authors.affiliation: Structural Glycobiology Section, CCR Nanobiology Program, Center for Cancer Research, National Cancer Institute at Frederick, Frederick, MD 21702, USA.
authors.forename: Boopathy
authors.initials: B
authors.lastname: Ramakrishnan
authors.forename: Pradman K
authors.initials: PK
authors.lastname: Qasba
mesh_terms: D000116:Acetylgalactosamine / Q000378:metabolism; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D002384:Catalysis; D004330:Drosophila / Q000201:enzymology; D019143:Evolution, Molecular; D005700:Galactosyltransferases / Q000737:chemistry; D006001:Glycoconjugates / Q000378:metabolism; D006801:Humans; D007448:Invertebrates / Q000201:enzymology / Q000378:metabolism*; D007768:Lactalbumin / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D008969:Molecular Sequence Data; D017350:N-Acetylgalactosaminyltransferases / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014530:Uridine Diphosphate / Q000378:metabolism; D014714:Vertebrates / Q000378:metabolism*
chemical_list: D006001:Glycoconjugates; D011134:Polysaccharides; D011994:Recombinant Proteins; D008345:Manganese; D014530:Uridine Diphosphate; D007768:Lactalbumin; D005700:Galactosyltransferases; D017350:N-Acetylgalactosaminyltransferases; C119701:beta-1,4-galactosyltransferase I; C023220:(N-acetylneuraminyl)-galactosylglucosylceramide N-acetylgalactosaminyltransferase; D000116:Acetylgalactosamine