Improved preparation and crystallization of 25 kDa human fibroblast growth factor-9.
We prepared 25 kDa human fibroblast growth factor-9 (hFGF-9 N33) on a large scale after overproduction in Escherichia coli MM294 (DE3)/pTG931. The purification was performed by a combination of hydrophobic chromatography and HPLC with an ion exchange column, a heparin affinity column and a gel filtration column. This improved procedure was rapid and simple, and the purified hFGF-9 N33 was found to be homogeneous as judged by various criteria, such as amino acid analysis, N-terminal amino acid sequence, C-terminal amino acid analysis and biological activity. Furthermore, as determined by low endotoxin and DNA content, the protein was of high purity. In addition, the hFGF-9 N33 prepared in the present study was easily crystallized by the vapour diffusion method.
Discovery Research Laboratory II, Pharmaceutical Discovery Research Division, Takeda Chemical Industries, Ltd., Wadai-10, Tsukuba, Ibaraki 300-4293, Japan.
N
N
Koyama
H
H
Ohmae
S
S
Tsuji
Y
Y
Tanaka
T
T
Kurokawa
O
O
Nishimura
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003460:Crystallization; D004058:Diffusion; D051525:Fibroblast Growth Factor 9; D005346:Fibroblast Growth Factors / Q000096:biosynthesis / Q000302:isolation & purification* / Q000502:physiology; D006493:Heparin; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight
C496459:FGF9 protein, human; C496460:Fgf9 protein, mouse; D051525:Fibroblast Growth Factor 9; D005346:Fibroblast Growth Factors; D006493:Heparin