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4YOL

Human fibroblast growth factor-1 C16S/A66C/C117A/P134A

Summary for 4YOL
Entry DOI10.2210/pdb4yol/pdb
Related1JQZ 1RG8 2AFG 3FJK 3HOM 4Q9G 4QAL
DescriptorFibroblast growth factor 1, CITRATE ANION, IMIDAZOLE, ... (4 entities in total)
Functional Keywordsfibroblast growth factor-1, cysteine-free mutant, fgf-1, intramolecular disulfide, protein binding
Biological sourceHomo sapiens (Human)
Total number of polymer chains2
Total formula weight33998.97
Authors
Xia, X.,Blaber, M. (deposition date: 2015-03-11, release date: 2016-03-16, Last modification date: 2024-10-16)
Primary citationXia, X.,Kumru, O.S.,Blaber, S.I.,Middaugh, C.R.,Li, L.,Ornitz, D.M.,Sutherland, M.A.,Tenorio, C.A.,Blaber, M.
Engineering a Cysteine-Free Form of Human Fibroblast Growth Factor-1 for "Second Generation" Therapeutic Application.
J.Pharm.Sci., 105:1444-1453, 2016
Cited by
PubMed Abstract: Human fibroblast growth factor-1 (FGF-1) has broad therapeutic potential in regenerative medicine but has undesirable biophysical properties of low thermostability and 3 buried cysteine (Cys) residues (at positions 16, 83, and 117) that interact to promote irreversible protein unfolding under oxidizing conditions. Mutational substitution of such Cys residues eliminates reactive buried thiols but cannot be accomplished simultaneously at all 3 positions without also introducing further substantial instability. The mutational introduction of a novel Cys residue (Ala66Cys) that forms a stabilizing disulfide bond (i.e., cystine) with one of the extant Cys residues (Cys83) effectively eliminates one Cys while increasing overall stability. This increase in stability offsets the associated instability of remaining Cys substitution mutations and permits production of a Cys-free form of FGF-1 (Cys16Ser/Ala66Cys/Cys117Ala) with only minor overall instability. The addition of a further stabilizing mutation (Pro134Ala) creates a Cys-free FGF-1 mutant with essentially wild-type biophysical properties. The elimination of buried free thiols in FGF-1 can substantially increase the protein half-life in cell culture. Here, we show that the effective cell survival/mitogenic functional activity of a fully Cys-free form is also substantially increased and is equivalent to wild-type FGF-1 formulated in the presence of heparin sulfate as a stabilizing agent. The results identify this Cys-free FGF-1 mutant as an advantageous "second generation" form of FGF-1 for therapeutic application.
PubMed: 27019961
DOI: 10.1016/j.xphs.2016.02.010
PDB entries with the same primary citation
Experimental method
X-RAY DIFFRACTION (1.97 Å)
Structure validation

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