Design of a thrombin resistant human acidic fibroblast growth factor (hFGF1) variant that exhibits enhanced cell proliferation activity.
Autor: | Kerr R; Department of Chemistry and Biochemistry, University of Arkansas, University of Arkansas, Fayetteville, AR, 72701, USA., Agrawal S; Department of Chemistry and Biochemistry, University of Arkansas, University of Arkansas, Fayetteville, AR, 72701, USA., Maity S; Department of Chemistry and Biochemistry, University of Arkansas, University of Arkansas, Fayetteville, AR, 72701, USA., Koppolu B; Joint Department of Biomedical Engineering, University of North Carolina-Chapel Hill & North Carolina State University, Raleigh, NC, 27695, USA., Jayanthi S; Department of Chemistry and Biochemistry, University of Arkansas, University of Arkansas, Fayetteville, AR, 72701, USA., Suresh Kumar G; Department of Chemistry and Biochemistry, University of Arkansas, University of Arkansas, Fayetteville, AR, 72701, USA., Gundampati RK; Department of Chemistry and Biochemistry, University of Arkansas, University of Arkansas, Fayetteville, AR, 72701, USA., McNabb DS; Department of Biological Sciences, University of Arkansas, University of Arkansas, Fayetteville, AR, 72701, USA., Zaharoff DA; Joint Department of Biomedical Engineering, University of North Carolina-Chapel Hill & North Carolina State University, Raleigh, NC, 27695, USA., Kumar TKS; Department of Chemistry and Biochemistry, University of Arkansas, University of Arkansas, Fayetteville, AR, 72701, USA. Electronic address: sthalla@uark.edu. |
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Jazyk: | angličtina |
Zdroj: | Biochemical and biophysical research communications [Biochem Biophys Res Commun] 2019 Oct 15; Vol. 518 (2), pp. 191-196. Date of Electronic Publication: 2019 Aug 13. |
DOI: | 10.1016/j.bbrc.2019.08.029 |
Abstrakt: | Acidic fibroblast growth factors (FGF1s) are heparin binding proteins that regulate a wide array of key cellular processes and are also candidates for promising biomedical applications. FGF1-based therapeutic applications are currently limited due to their inherent thermal instability and susceptibility to proteases. Using a wide range of biophysical and biochemical techniques, we demonstrate that reversal of charge on a well-conserved positively charged amino acid, R136, in the heparin binding pocket drastically increases the resistance to proteases, thermal stability, and cell proliferation activity of the human acidic fibroblast growth factor (hFGF1). Two-dimensional NMR data suggest that the single point mutations at position-136 (R136G, R136L, R136Q, R136K, and R136E) did not perturb the backbone folding of hFGF1. Results of the differential scanning calorimetry experiments show that of all the designed R136 mutations only the charge reversal mutation, R136E, significantly increases (ΔT (Copyright © 2019 Elsevier Inc. All rights reserved.) |
Databáze: | MEDLINE |
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