Regulating cell surface glycosylation by small molecule control of enzyme localization.
Cell surface carbohydrates mediate interactions between the cell and its environment. Glycosyltransferases responsible for synthesis of cell surface oligosaccharides are therefore essential administrators of cellular communication. These enzymes often comprise large families. Redundancy of related family members and embryonic lethality both complicate genetic methods for deconvoluting functions of glycosyltransferases. We report a chemical method in which the activity of an individual glycosyltransferase is controlled by a small molecule. The approach exploits the requirement of Golgi localization, a common feature of glycosyltransferase superfamily members. In our approach, the glycosyltransferase is separated into two domains, one that determines localization and one responsible for catalysis. Control of enzyme activity is achieved using a small molecule to regulate association of the two domains. We used this method to regulate production of sialyl Lewis x by alpha1,3-fucosyltransferase VII in living cells.
Kohler
Jennifer J
JJ
Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720, USA.
Bertozzi
Carolyn R
CR
D000818:Animals; D016466:CHO Cells; D002384:Catalysis; D020134:Catalytic Domain; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism*; D006224:Cricetinae; D019281:Dimerization; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D006031:Glycosylation / Q000187:drug effects; D016695:Glycosyltransferases / Q000737:chemistry / Q000378:metabolism*; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020123:Sirolimus / Q000378:metabolism / Q000494:pharmacology*; D022021:Tacrolimus Binding Proteins / Q000235:genetics / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D016695:Glycosyltransferases; D022021:Tacrolimus Binding Proteins; D020123:Sirolimus