Construction of a Sensitive Enzyme Immunoassay for Human Fibroblast Growth Factor 9

Matsumoto-Yoshitomi, Sumie; Kuroshima, Ken-Ichi; Nomura, Chisako; Habashita, Junko; Kurokawa, Tsutomu

doi:10.1089/hyb.1996.15.299

Cited by 5 publications

(2 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Structure Determination-The first 33 N-terminal residues of FGF9 were deleted because they are implicated only in secretion and not FGFR binding (17,26). Truncated FGF9 (residues 35-208) was expressed in E. coli and purified to homogeneity (see "Experimental Procedures").…”

Section: Resultsmentioning

confidence: 99%

Crystal Structure of Fibroblast Growth Factor 9 Reveals Regions Implicated in Dimerization and Autoinhibition

Plotnikov¹,

Eliseenkova²,

Ibrahimi³

et al. 2001

Journal of Biological Chemistry

View full text Add to dashboard Cite

Fibroblast growth factors (FGFs) constitute a large family of heparin-binding growth factors with diverse biological activities. FGF9 was originally described as glia-activating factor and is expressed in the nervous system as a potent mitogen for glia cells. Unlike most FGFs, FGF9 forms dimers in solution with a K d of 680 nM. To elucidate the molecular mechanism of FGF9 dimerization, the crystal structure of FGF9 was determined at 2.2 Å resolution. FGF9 adopts a ␤-trefoil fold similar to other FGFs. However, unlike other FGFs, the N-and C-terminal regions outside the ␤-trefoil core in FGF9 are ordered and involved in the formation of a 2-fold crystallographic dimer. A significant surface area (>2000 Å 2 ) is buried in the dimer interface that occludes a major receptor binding site of FGF9. Thus, we propose an autoinhibitory mechanism for FGF9 that is dependent on sequences outside of the ␤-trefoil core. Moreover, a model is presented providing a molecular basis for the preferential affinity of FGF9 toward FGFR3.The mammalian fibroblast growth factor (FGF) 1 family contains at least 22 distinct polypeptides (FGF1-FGF22) that are expressed in a specific spatial and temporal pattern (1-5). FGFs play important roles in numerous physiological and pathological processes (1-5). Members of the FGF family share between 10 and 55% sequence identity (6). Crystal structures of FGF1 (7), FGF2 (7-9), and FGF7 (10), have revealed a common core FGF structure consisting of three copies of a four-stranded ␤-sheet, known as a ␤-trefoil fold. All FGFs contain N-and C-terminal segments outside the ␤-trefoil core. However, the length of these segments, especially the C termini, varies greatly among different FGFs. In FGF1, FGF2, FGF4, and FGF7, the end of polypeptide chain virtually coincides with the end of the ␤-trefoil fold. Consequently, these FGFs have extremely short C-terminal segments. In contrast, other FGFs, such as FGF3, FGF5, and FGF8, have long C-terminal extensions. The functional relevance of these N-and C-terminal extensions remains uncharacterized. FGF9 (glia-activating factor) was purified as a heparin-binding, secreted glycoprotein from cultured human glioma cell line NMC-G1 (11). FGF9 shows 30% sequence identity with the prototypical FGF family members, FGF1 and FGF2 (12). Purified FGF9 is mitogenic for many types of cultured cells including glia cells, oligodendrocyte type 2 astrocyte progenitor cells, smooth muscle cells, pheochromocytoma PC12 cells, and BALB/3T3 fibroblasts (11). Because FGF9, unlike FGF1 and FGF2, has no effect on human umbilical vein endothelial cells, it is suggested that FGF9 may have a unique receptor specificity (11). In fact, biochemical studies performed utilizing various soluble FGFR-alkaline phosphatase fusion proteins and genetically engineered cells expressing different full-length FGFRs have demonstrated that FGF9 binds preferentially to the IIIc form of FGFR3 (13-15).FGF9, like prototypical FGFs, does not have a typical secretory signal peptide. Yet, it is still glycosylated and...

show abstract

Section: Resultsmentioning

confidence: 99%

Crystal Structure of Fibroblast Growth Factor 9 Reveals Regions Implicated in Dimerization and Autoinhibition

Plotnikov¹,

Eliseenkova²,

Ibrahimi³

et al. 2001

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Immünogen olarak recombinant DNA'lar kullanılmaktadır. Monoclonal bir antibody olan Anti-FGF-9 uygun dozlarda verildiği takdirde in vivo ve in vitro şartlarda FGF-9'u bloke ederek onun biyolojik aktivitesini ortadan kaldırmaktadır (12).…”

Section: Introductionunclassified

The Effects Of Fgf-9 On In Vitro Embryonic Development

Tekinarslan¹,

Unur²,

Ülger³

et al. 2009

TUTFD

View full text Add to dashboard Cite

Autocrine transformation by fibroblast growth factor 9 (FGF-9) and its possible participation in human oncogenesis

et al. 1997

Self Cite

View full text Add to dashboard Cite

Transfection of human fibroblast growth factor 9 (FGF-9) cDNA into mouse BALB/c 3T3 clone A31 cells led to morphological transformation of the cells and foci formation 4 weeks later. Isolated transformants had a higher saturation density than parental A31 cells, could grow in soft agar, and secreted FGF-9 into the culture supernatant. The introduction of FGF-9 N33 cDNA, which encodes a truncated protein that has 33 N-terminal amino acids deleted and has the same mitogenic potency as FGF-9, failed to lead to foci formation. Although FGF-9 is a secretory protein, it does not have a typical secretory signal sequence, and the secreted protein retains the full sequence coded in the cDNA except for the initiating methionine. The produced FGF-9 N33 was not secreted and remained within the cell. It is possible that FGF-9 has an uncleavable signal sequence within the first 33 N-terminal amino acids. All of the phenotypes acquired by transformation could be arrested by treatment with a neutralizing anti-human FGF-9 monoclonal antibody (MAb) 150-59. Additionally, transformants formed tumors in nude mice. Injection of MAb 150-59 suppressed tumor formation in nude mice and caused existing tumors to regress. Our results suggest that the cellular transformation mediated by FGF-9 is produced by autocrine stimulation. We have detected FGF-9 production in the human tumor cell lines glioma NMC-G1, from which FGF-9 was originally purified, and stomach carcinoma AZ-521. The growth of NMC-G1 was not affected by MAb 150-59, but that of AZ-521 was arrested by MAb 150-59 in the presence of heparin. Moreover, the growth of the AZ-521 cell tumor in nude mice could be partially arrested by antibody treatment. The possibility of a participation of FGF-9 in the formation of human tumors is suggested. Int. J.

show abstract

Construction of a Sensitive Enzyme Immunoassay for Human Fibroblast Growth Factor 9

Cited by 5 publications

References 11 publications

Crystal Structure of Fibroblast Growth Factor 9 Reveals Regions Implicated in Dimerization and Autoinhibition

Crystal Structure of Fibroblast Growth Factor 9 Reveals Regions Implicated in Dimerization and Autoinhibition

The Effects Of Fgf-9 On In Vitro Embryonic Development

Autocrine transformation by fibroblast growth factor 9 (FGF-9) and its possible participation in human oncogenesis

Contact Info

Product

Resources

About