A1 Refereed original research article in a scientific journal
Expression and glycosylation studies of human FGF receptor 4
Authors: Tuominen H, Heikinheimo P, Loo B-M, Kataja K, Oker-Blom C, Uutela M, Jalkanen M, Goldman A
Publisher: ACADEMIC PRESS INC
Publication year: 2001
Journal:: Protein Expression and Purification
Journal name in source: PROTEIN EXPRESSION AND PURIFICATION
Journal acronym: PROTEIN EXPRES PURIF
Volume: 21
Issue: 2
First page : 275
Last page: 285
Number of pages: 11
ISSN: 1046-5928
DOI: https://doi.org/10.1006/prep.2000.1375
Abstract
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.
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.
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