Expression and glycosylation studies of human FGF receptor 4

被引：10

作者：

Tuominen, H

Heikinheimo, P

Loo, BM

Kataja, K

Oker-Blom, C

Uutela, M

Jalkanen, M

Goldman, A

机构：

[1] Univ Helsinki, Inst Biotechnol, Mol Canc Biol Lab, FIN-00014 Helsinki, Finland

[2] Univ Turku, Turku Ctr Biotechnol, FIN-20521 Turku, Finland

[3] Abo Akad Univ, FIN-20521 Turku, Finland

[4] VTT Biotechnol & Food Res, FIN-02044 VTT, Finland

[5] Univ Jyvaskyla, Dept Biol & Environm Sci, Div Biotechnol, FIN-40351 Jyvaskyla, Finland

[6] BioTie Therapies Ltd, FIN-20520 Turku, Finland

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2001年 / 21卷 / 02期

基金：

芬兰科学院;

关键词：

glycosylation; expression; human FGFR4; purification; refolding;

D O I：

10.1006/prep.2000.1375

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 FCFR4 extracellular domain (FGFR4(ed)) for structural studies. We show that baculovirus-insect cell-expressed FGFR4(ed) 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 FGFR4(ed), but was still heterogeneous. Large amounts of FGFRA(ed) have been produced into inclusion bodies in Escherichia coli and refolded at least partly correctly but the refolded E. coli-produced FGFR4(ed) still aggregates. (C) 2001 Academic Press.

引用

页码：275 / 285

页数：11

共 28 条

[1] EXPRESSION OF A DOMINANT NEGATIVE MUTANT OF THE FGF RECEPTOR DISRUPTS MESODERM FORMATION IN XENOPUS EMBRYOS [J].