Deletion of the cytoplasmic domain of human alpha3/4 fucosyltransferase III causes the shift of the enzyme to early Golgi compartments.
The transmembrane domain (TM) and flanking regions of glycosyltransferases (GTs) have been implicated in the localization of these proteins in the Golgi apparatus (GA). alpha3/4 Fucosyltransferase III (FT3wt) (EC 2.4.1.65) is localized in the trans-Golgi and trans-Golgi network (TGN) of baby hamster kidney (BHK) cells and synthesizes Lewis determinants associated with cell adhesion events. We have evaluated the effect of removing the cytosolic domain on the localization of the enzyme and its capacity for synthesizing the Lewis A (Le A) determinant. The mutant where the cytoplasmic domain (Asp-2 to Trp-13) of FT3wt has been deleted (FT3dc) was localized in the Golgi but it was shifted to earlier compartments than FT3wt. The mutant was not detected on the plasma membrane (PM) and glycosylation analysis indicated that FT3dc was transported beyond the endoplasmic reticulum (ER) since complex type glycosylation was observed. Cells expressing FT3dc showed a significantly lower efficiency to synthesize Le A when compared with cells expressing FT3wt, in vivo. This reduction was not due to lower specific activity because both enzyme forms had a similar specific activity in vitro. Therefore, removal of FT3 cytosolic tail caused a shift in enzyme distribution to earlier Golgi compartments concomitant to the decrease of its biosynthetic capacity.
Sousa
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VL
Instituto de Tecnologia Química e Biológica, Avenida da República, Apartado 127, 2781-901 Oeiras, Portugal.
Brito
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C
Costa
J
J
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D006224:Cricetinae; D003593:Cytoplasm; D005647:Fucosyltransferases / Q000737:chemistry / Q000378:metabolism*; D006031:Glycosylation; D006056:Golgi Apparatus / Q000201:enzymology*; D006801:Humans; D007668:Kidney / Q000196:embryology; D007983:Lewis Blood Group Antigens / Q000378:metabolism; D009154:Mutation; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D017384:Sequence Deletion; D021601:trans-Golgi Network
D007983:Lewis Blood Group Antigens; D005647:Fucosyltransferases; C019818:3-galactosyl-N-acetylglucosaminide 4-alpha-L-fucosyltransferase