Catalytic and glycan-binding abilities of ppGalNAc-T2 are regulated by acetylation

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Natacha Zlocowski, Victor G Sendra, Virginia Lorenz, Marcos A Villarreal, Alberto Jorge, Yolanda Núñez, Eric P Bennett, Henrik Clausen, Gustavo A Nores, Fernando J Irazoqui

Post-translational acetylation is an important molecular regulatory mechanism affecting the biological activity of proteins. Polypeptide GalNAc transferases (ppGalNAc-Ts) are a family of enzymes that catalyze initiation of mucin-type O-glycosylation. All ppGalNAc-Ts in mammals are type II transmembrane proteins having a Golgi lumenal region that contains a catalytic domain with glycosyltransferase activity, and a C-terminal R-type ("ricin-like") lectin domain. We investigated the effect of acetylation on catalytic activity of glycosyltransferase, and on fine carbohydrate-binding specificity of the R-type lectin domain of ppGalNAc-T2. Acetylation effect on ppGalNAc-T2 biological activity in vitro was studied using a purified human recombinant ppGalNAc-T2. Mass spectrometric analysis of acetylated ppGalNAc-T2 revealed seven acetylated amino acids (K103, S109, K111, K363, S373, K521, and S529); the first five are located in the catalytic domain. Specific glycosyltransferase activity of ppGalNAc-T2 was reduced 95% by acetylation. The last two amino acids, K521 and S529, are located in the lectin domain, and their acetylation results in alteration of the carbohydrate-binding ability of ppGalNAc-T2. Direct binding assays showed that acetylation of ppGalNAc-T2 enhances the recognition to aGalNAc residue of MUC1aGalNAc, while competitive assays showed that acetylation modifies the fine GalNAc-binding form of the lectin domain. Taken together, these findings clearly indicate that biological activity (catalytic capacity and glycan-binding ability) of ppGalNAc-T2 is regulated by acetylation.
TidsskriftBiochemical and Biophysical Research Communications
Udgave nummer1
Sider (fra-til)140-5
Antal sider6
StatusUdgivet - 2011

ID: 38489984