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Pituitary fibroblast growth factors: partial purification and characterization
Summary
Researchers separated acidic and basic fibroblast growth factor (FGF) using isoelectric focusing. Acidic FGF showed unique stability and precipitation properties, demonstrating potent DNA synthesis initiation activity.
Area of Science:
- Biochemistry
- Cell Biology
- Molecular Biology
Background:
- Fibroblast growth factors (FGFs) are crucial for cell proliferation and differentiation.
- Separating acidic and basic FGF from biological extracts presents challenges due to potential interactions.
Purpose of the Study:
- To develop a method for separating acidic and basic FGF activities from bovine pituitary extracts.
- To characterize the distinct properties and biological activity of acidic and basic FGF.
Main Methods:
- Isoelectric focusing (IEF) was employed to separate FGF activities based on their isoelectric points (pI).
- Chromatography on CM-Sephadex was initially attempted but proved ineffective for separation.
- Fibroblast growth factor activity was quantified by measuring DNA synthesis initiation in Swiss mouse 3T3 fibroblasts.
Main Results:
- Isoelectric focusing successfully separated acidic FGF (pI 4-5) from basic FGF (pI 8-9).
- Acidic FGF exhibited stability in urea and beta-mercaptoethanol, unlike basic FGF, which was inactivated.
- Acidic FGF precipitated at pH 4.5 and redissolved at pH 8, while basic FGF remained soluble and stable at pH 4.5.
- Acidic FGF demonstrated potent DNA synthesis initiation activity, comparable to established basic FGF preparations.
Conclusions:
- Isoelectric focusing is an effective technique for resolving acidic and basic FGF activities.
- Acidic and basic FGF possess distinct biochemical properties influencing their stability and solubility.
- Acidic FGF is a potent stimulator of DNA synthesis, highlighting its biological significance.