Production and characterization of the extracellular domain of recombinant human fibroblast growth factor receptor 4.
Among the members of the fibroblast growth factor receptor family the FGFR4 has demonstrated strong dependence on heparin-like material for its activation by fibroblast growth factors. We have produced and characterized a recombinant human FGFR4 extracellular domain (FGFR4ed), in order to study its biochemical properties in isolated conditions. The FGFR4ed was expressed in an insect cell system and purified from the culture medium by Ni(2+)-affinity and gel filtration chromatography. Pure FGFR4ed was tested for FGF- and heparin-binding by covalent crosslinking experiments and by biosensor analysis. In solution, FGFR4ed formed complexes with acidic FGF (FGF-1) and basic FGF (FGF-2), both in the presence and absence of heparin. Immobilized FGFR4 also bound FGF-8 besides FGF-1 and FGF-2. Furthermore, heparin alone induced receptor oligomerization on the surface of the receptor coupled chip. Thus, the recombinant FGFR4ed revealed properties described for the cellular form of this receptor and can be used for interaction studies.
Loo
B B
BB
Turku Centre for Biotechnology, Abo Akademi University and University of Turku, FIN-20520, Turku, Finland. bmloo@btk.utu.fi
Darwish
K K
KK
Vainikka
S S
SS
Saarikettu
J J
JJ
Vihko
P P
PP
Hermonen
J J
JJ
Goldman
A A
AA
Alitalo
K K
KK
Jalkanen
M M
MM
D000818:Animals; D015374:Biosensing Techniques; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D019281:Dimerization; D016220:Fibroblast Growth Factor 1; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D051524:Fibroblast Growth Factor 8; D005346:Fibroblast Growth Factors / Q000378:metabolism; D006493:Heparin / Q000378:metabolism; D006639:Histidine; D006801:Humans; D007313:Insecta / Q000166:cytology; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D051499:Receptor, Fibroblast Growth Factor, Type 4; D017468:Receptors, Fibroblast Growth Factor / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
C496456:FGF8 protein, human; D010455:Peptides; D017468:Receptors, Fibroblast Growth Factor; D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; D016220:Fibroblast Growth Factor 1; D051524:Fibroblast Growth Factor 8; C033223:polyhistidine; D006639:Histidine; D005346:Fibroblast Growth Factors; D006493:Heparin; C496371:FGFR4 protein, human; D051499:Receptor, Fibroblast Growth Factor, Type 4