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Soluble kit receptor in human serum
J Wypych1, L G Bennett, M G Schwartz
1Amgen, Inc, Amgen Center, Thousand Oaks, CA 91320-1789.
Blood
|January 1, 1995
Summary
Researchers developed a new assay to measure soluble Kit in human serum, finding high levels that do not correlate with age or sex. This soluble Kit may regulate stem cell factor (SCF) activity in vivo.
Area of Science:
- Molecular Biology
- Immunology
- Biochemistry
Background:
- c-kit encodes the transmembrane receptor tyrosine kinase (Kit) for stem cell factor (SCF).
- Soluble forms of receptor tyrosine kinases can influence ligand binding and signaling.
Purpose of the Study:
- To develop and validate an assay for soluble human Kit in serum.
- To characterize the biochemical properties and potential function of soluble Kit.
Main Methods:
- Enzyme-linked immunosorbent assay (ELISA) for soluble Kit quantification.
- Immunoaffinity chromatography for purification.
- Immunoblotting, N-terminal sequencing, and glycosylation analysis.
- In vitro assay to assess inhibition of SCF binding.
Main Results:
- High levels of soluble Kit were detected in normal human serum (mean 324 +/- 105 ng/mL).
- Soluble Kit levels showed no correlation with donor sex or age.
- Purified soluble Kit is glycosylated and shares characteristics with recombinant soluble Kit.
- Serum-derived soluble Kit inhibited SCF binding to membrane-bound receptors.
Conclusions:
- Soluble Kit is present at significant levels in human serum.
- The characterized soluble Kit is biochemically similar to the extracellular domain of the membrane-bound receptor.
- Soluble Kit has the potential to modulate SCF activity in vivo.