Visualizing specific protein glycoforms by transmembrane fluorescence resonance energy transfer.
Analyses of mice lacking glycosyltransferase have suggested that their pathological phenotypes are not attributable to the overall change of the sugar modification, but instead the result of changes of the glycan structures on a specific 'target' glycoprotein. Therefore, detecting or monitoring the glycosylation status of a specific protein in living cells is important, but no such methods are currently available. Here we demonstrate the detection of glycoforms of a specific glycoprotein using the fluorescence resonance energy transfer technique. Using model proteins, we detect characteristic fluorescence resonance energy transfer signals from the specific glycoform-bearing target glycoprotein. We also show that, upon insulin removal, sialylated glycoforms of green fluorescent protein-tagged GLUT4 seem to be internalized more slowly than non-sialylated GLUT4. This novel analytical imaging tool allows studying the roles of specific glycan modifications of a protein of interest.
Glycometabolome Team, RIKEN Advanced Science Institute, Wako, Saitama 351-0198, Japan.
Yoshimi
Y
Haga
Kumiko
K
Ishii
Kayo
K
Hibino
Yasushi
Y
Sako
Yukishige
Y
Ito
Naoyuki
N
Taniguchi
Tadashi
T
Suzuki
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D051275:Glucose Transporter Type 4 / Q000737:chemistry / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*
D051275:Glucose Transporter Type 4; D006023:Glycoproteins; D011506:Proteins; D049452:Green Fluorescent Proteins