Formation of insoluble oligomers correlates with ST6Gal I stable localization in the golgi.
The ST6Gal I is a sialyltransferase that functions in the late Golgi to modify the N-linked oligosaccharides of glycoproteins. The ST6Gal I is expressed as two isoforms with a single amino acid difference in their catalytic domains. The STcys isoform is stably retained in the cell and is predominantly found in the Golgi, whereas the STtyr isoform is only transiently localized in the Golgi and is cleaved and secreted from a post-Golgi compartment. These two ST6Gal I isoforms were used to explore the role of the bilayer thickness mechanism and oligomerization in Golgi localization. Analysis of STcys and STtyr proteins with longer transmembrane regions suggested that the bilayer thickness mechanism is not the predominant mechanism used for ST6Gal I Golgi localization. In contrast, the formation and quantity of Triton X-100-insoluble oligomers was correlated with the stable or transient localization of the ST6Gal I isoforms in the Golgi. Nearly 100% of the STcys and only 13% of the STtyr were found as Triton-insoluble oligomers when Golgi membranes of COS-1 cells expressing these proteins were solubilized at pH 6.3, the pH of the late Golgi. In contrast, both proteins were found in the soluble fraction when these membranes were solubilized at pH 8.0. Analysis of other mutants suggested that a conformational change in the catalytic domain rather than increased disulfide bond-based cross-linking is the basis for the increased ability of STcys protein to form oligomers and the stable localization of STcys protein in the Golgi.
Chen
C
C
Department of Biochemistry, University of Illinois at Chicago, College of Medicine, Chicago, Illinois 60612, USA.
Ma
J
J
Lazic
A
A
Backovic
M
M
Colley
K J
KJ
D000818:Animals; D001692:Biological Transport; D019556:COS Cells; D002451:Cell Compartmentation; D019281:Dimerization; D006056:Golgi Apparatus / Q000201:enzymology*; D011487:Protein Conformation; D012799:Sialyltransferases / Q000737:chemistry* / Q000378:metabolism*; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D012799:Sialyltransferases; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase