TY - JOUR
T1 - Expression and glycosylation studies of human FGF receptor 4
AU - Tuominen, Heidi
AU - Heikinheimo, Pirkko
AU - Loo, Britt-Marie
AU - Kataja, Kari
AU - Oker-Blom, Christian
AU - Uutela, Marko
AU - Jalkanen, Markkku
AU - Goldman, Adrian
PY - 2001
Y1 - 2001
N2 - Fibroblast growth factor receptor subtype 4 (FGFR4) has been shown to have special activation properties and just one splicing form, unlike the other FGFRs. FGFR4 overexpression is correlated with breast cancer and therefore FGFR4 is a target for drug design. Our aim is to overexpress high amounts of homogeneous FGFR4 extracellular domain (FGFR4ed) for structural studies. We show that baculovirus-insect cell-expressed FGFR4ed is glycosylated on three (N88, N234, and N266) of the six possible N-glycosylation sites but is not O-glycosylated. The deglycosylated triple mutant was expressed and had binding properties similar to those of glycosylated FGFR4ed, but was still heterogeneous. Large amounts of FGFR4ed have been produced into inclusion bodies in Escherichia coli and refolded at least partly correctly but the refolded E. coli-produced FGFR4ed still aggregates.
AB - Fibroblast growth factor receptor subtype 4 (FGFR4) has been shown to have special activation properties and just one splicing form, unlike the other FGFRs. FGFR4 overexpression is correlated with breast cancer and therefore FGFR4 is a target for drug design. Our aim is to overexpress high amounts of homogeneous FGFR4 extracellular domain (FGFR4ed) for structural studies. We show that baculovirus-insect cell-expressed FGFR4ed is glycosylated on three (N88, N234, and N266) of the six possible N-glycosylation sites but is not O-glycosylated. The deglycosylated triple mutant was expressed and had binding properties similar to those of glycosylated FGFR4ed, but was still heterogeneous. Large amounts of FGFR4ed have been produced into inclusion bodies in Escherichia coli and refolded at least partly correctly but the refolded E. coli-produced FGFR4ed still aggregates.
U2 - 10.1006/prep.2000.1375
DO - 10.1006/prep.2000.1375
M3 - Article
SN - 1046-5928
VL - 21
SP - 275
EP - 285
JO - Protein Expression and Purification
JF - Protein Expression and Purification
IS - 2
ER -
