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Purification of a human urinary colony-stimulating factor
Journal of Cellular Biochemistry
|January 1, 1983
Summary
Colony-stimulating factor (CSF) was isolated from anemic patient urine, revealing a purified protein essential for granulocyte and macrophage colony formation. The active form of this urinary CSF is a dimer, distinct from mouse L-cell CSF.
Area of Science:
- Biochemistry
- Hematology
- Protein Chemistry
Background:
- Colony-stimulating factor (CSF) is crucial for the in vitro development of granulocytes and macrophages.
- Understanding CSF's properties is vital for research into hematopoiesis and related disorders.
Purpose of the Study:
- To isolate and characterize colony-stimulating factor (CSF) from the urine of anemic patients.
- To determine the molecular properties and biological activity of purified urinary CSF.
Main Methods:
- A seven-step isolation procedure was employed to purify CSF from urine.
- Sodium dodecyl sulfate (SDS) gel electrophoresis and mercaptoethanol reduction were used to assess molecular weight.
- Mouse bone marrow cell culture assays were performed to evaluate colony formation stimulation.
Main Results:
- Homogeneous urinary CSF was obtained with high specific activity (1.9 X 10(8) U/AU), representing a 25,330-fold purification.
- The purified CSF exists as a dimer (46,000 MW) that dissociates into a 23,000 MW component upon reduction; only the dimer is biologically active.
- Urinary CSF specifically stimulated macrophage colony formation and showed distinct binding characteristics with an antibody against mouse L-cell CSF.
Conclusions:
- The study successfully isolated and purified biologically active CSF from human urine.
- Urinary CSF is a dimeric glycoprotein essential for macrophage colony formation, with structural and functional differences compared to mouse L-cell CSF.