title: An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2.
abstract: Polypeptide GalNAc-transferases (ppGalNAc-Ts) are a family of enzymes that catalyze the initiation of mucin-type O-glycosylation. All ppGalNAc-T family members contain a common (QXW)3 motif, which is present in the R-type lectin group. The acetylation site K521 is part of the QKW motif of β-trefoil in the lectin domain of ppGalNAc-T2. We used a combination of acetylation and site-directed mutagenesis approaches to examine the functional role of K521 in ppGalNAc-T2. Binding assays of non-acetylated and acetylated forms of the mutant ppGalNAc-T2K521Q to various naked and αGalNAc-glycosylated mucin peptides indicated that the degree of interaction of lectin domain with αGalNAc depends on the peptide sequence of mucin. Studies of the inhibitory effect of various carbohydrates on the interactions of ppGalNAc-T2 with MUC1αGalNAc indicate that point K521Q mutation enhance the carbohydrate specificity of lectin domain for αGalNAc. K521Q mutation resulted in an enzyme activity lower than that of the wild-type ppGalNAc-T2, similar to the acetylation of ppGalNAc-T2. We conclude that an acetylation site in the QKW motif of the lectin domain modulates carbohydrate recognition specificity and catalytic activity of ppGalNAc-T2 for partially preglycosylated acceptors and a certain naked peptide. Posttranslational modifications of ppGalNAc-Ts, such as acetylation, may play key roles in modulating the functions of the R-type lectin domains in cellular homeostasis.
authors.affiliation: Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC, UNC– CONICET), Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Cordoba, 5000 Córdoba, Argentina.
authors.forename: Natacha
authors.initials: N
authors.lastname: Zlocowski
authors.forename: Virginia
authors.initials: V
authors.lastname: Lorenz
authors.forename: Eric P
authors.initials: EP
authors.lastname: Bennett
authors.forename: Henrik
authors.initials: H
authors.lastname: Clausen
authors.forename: Gustavo A
authors.initials: GA
authors.lastname: Nores
authors.forename: Fernando J
authors.initials: FJ
authors.lastname: Irazoqui
mesh_terms: D000107:Acetylation; D006801:Humans; D037102:Lectins / Q000737:chemistry / Q000378:metabolism*; D017350:N-Acetylgalactosaminyltransferases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000097763:Polypeptide N-acetylgalactosaminyltransferase
chemical_list: D037102:Lectins; D017350:N-Acetylgalactosaminyltransferases; D000097763:Polypeptide N-acetylgalactosaminyltransferase