Probing the role of proline -135 on the structure, stability, and cell proliferation activity of human acidic fibroblast growth factor.


Abstract

Human acidic fibroblast growth factor 1 (hFGF1) is a protein intricately involved in cell growth and tissue repair. In this study, we investigate the effect(s) of understanding the role of a conserved proline (P135), located in the heparin binding pocket, on the structure, stability, heparin binding affinity, and cell proliferation activity of hFGF1. Substitution of proline-135 with a positively charged lysine (P135K) resulted in partial destabilization of the protein; however, the overall structural integrity of the protein was maintained upon substitution of proline-135 with either a negative charge (P135E) or a polar amino acid (P135Q). Interestingly, upon heparin binding, an increase in thermal stability equivalent to that of wt-hFGF1 was observed when P135 was replaced with a positive (P135K) or a negative charge (P135E), or with a polar amino acid (P135Q). Surprisingly, introduction of negative charge in the heparin-binding pocket at position 135 (P135E) increased hFGF1's affinity for heparin by 3-fold, while the P135K mutation, did not alter the heparin-binding affinity. However, the enhanced heparin-binding affinity of mutant P135E did not translate to an increase in cell proliferation activity. Interestingly, the P135K and P135E double mutations, P135K/R136E and P135/R136E, reduced the heparin binding affinity by ∼3-fold. Furthermore, the cell proliferation activity was increased when the charge reversal mutation R136E was paired with both P135E (P135E/R136E) and P135K (P135K/R136E). Overall, the results of this study suggest that while heparin is useful for stabilizing hFGF1 on the cell surface, this interaction is not mandatory for activation of the FGF receptor.

Submission Details

ID: SgQVErXY

Submitter: Connie Wang

Submission Date: Sept. 28, 2018, 4:24 p.m.

Version: 1

Publication Details
Davis JE;Alghanmi A;Gundampati RK;Jayanthi S;Fields E;Armstrong M;Weidling V;Shah V;Agrawal S;Koppolu BP;Zaharoff DA;Kumar TKS,Arch Biochem Biophys (2018) Probing the role of proline -135 on the structure, stability, and cell proliferation activity of human acidic fibroblast growth factor. PMID:30031837
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