Thr-114 is an important functional residue of fibroblast growth factor 10 identified by structure-based mutational analysis.
Fibroblast growth factor 10 (FGF10) plays important roles in vertebrate limb development, lung branching morphogenesis, and epidermis regeneration. The receptor (FGFR2b) binding specificity is an essential element in regulating the diverse functions of FGF10. Analyzing the FGF10:FGFR2b complex we found that Thr-114 in beta4 of FGF10 could form specific interactions with D3 of FGFR2b. To investigate the role of Thr-114 played on functions of FGF10, two mutants of FGF10 were constructed, named TA (Thr-114-->Ala) and TR (Thr-114-->Arg), respectively. The biological activity assays showed that the receptor-binding affinity, the stimulating growth effect on rat tracheal epithelium (RTE) cells, and the inducing ability in receptor phosphorylation of both mutants were decreased, which were consistent with the interaction analysis of the TA:FGFR2b and TR:FGFR2b complexes. These results suggested that Thr-114 is a crucial functional residue for FGF10, and mutating Thr-114 to Ala or Arg would lead to great decrease in receptor-binding affinity and biological activity of FGF10.
Wang
Jin-feng
JF
Institute of Basic Medical Sciences, Beijing, China.
Cai
Xin
X
Zou
Min-ji
MJ
Wang
Yuan-yuan
YY
Wang
Jia-xi
JX
Xu
Dong-gang
DG
D000818:Animals; D004252:DNA Mutational Analysis; D051526:Fibroblast Growth Factor 10 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D041681:NIH 3T3 Cells; D017354:Point Mutation; D011487:Protein Conformation; D051381:Rats; D051497:Receptor, Fibroblast Growth Factor, Type 2 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013912:Threonine / Q000235:genetics / Q000378:metabolism*
C496463:FGF10 protein, human; D051526:Fibroblast Growth Factor 10; D011994:Recombinant Proteins; D013912:Threonine; D051497:Receptor, Fibroblast Growth Factor, Type 2