Functional Analysis of Glycosaminoglycans in the complex formation of FGFs and their receptors
| Project/Area Number |
16570128
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Functional biochemistry
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| Research Institution | National Institute of Advanced Industrial Science and Technology |
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Principal Investigator |
ASADA Masahiro National Institute of Advanced Industrial Science and Technology, Signaling Molecules Research Laboratory, Senior Research Scientist, シグナル分子研究ラボ, 主任研究員 (30344120)
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| Co-Investigator(Kenkyū-buntansha) |
IMAMURA Toru National Institute of Advanced Industrial Science and Technology, Signaling Molecules Research Laboratory, Director, シグナル分子研究ラボ, 研究ラボ長 (80356518)
SUZUKI Masashi National Institute of Advanced Industrial Science and Technology, Signaling Molecules Research Laboratory, Senior Research Scientist, シグナル分子研究ラボ, 主任研究員 (70192622)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2005: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2004: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | heparin-binding growth factor / glycosaminoglycan / recombinant protein / fibroblast growth factor |
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| Research Abstract |
Twenty-two ligands of fibroblast growth factor (FGF) family and seven major variants of their receptor (FGFR) have been identified. Earlier studies revealed that the ligand and the receptor form biologically active signaling complex on the cell surface, only when heparn sulfate or heparin is involved. Recently some FGFs were reported to bind chondroitin sulfate as well. To address the potential roles of various glycosaminoglycans (GAG) in the regulation of FGF activity and its signaling, we investigated the affinity of FGFs to various GAGs and involvement of GAGs on the formation of FGF : FGFR complex.
The cDNAs for FGFs and FGFRs were prepared, and the proteins were expressed by E. coli with FLAG-tag and by COS-1 cells as IgG(Fc)-fusion proteins, respectively. FGFs loaded on the immobilized GAG columns were eluted with a buffer containing increasing concentration of NaCl and their affinity for the respective GAG were determined. And FGFs were incubated with immobilized receptors in the presence or absence of various GAGs, and the bound ligands were detected by ELISA.
Using these assay systems, we found that most FGFs bound tightly to heparin as reported and also found additional binding combinations of FGF and GAG. We also detected the requirement of heparin in the formation of the FGF : FGFR complex and further identified additional combinations of the specific GAG : FGF : FGFR complex. These interaction may suggest biological importance of these complex formations in growth factors' activity.
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Report
(4 results)
Research Products
(6 results)
