Molecular weights of glycosylated and nonglycosylated forms of recombinant human stem cell factor determined by low-angle laser light scattering.
The molecular weight of recombinant human stem cell factor (SCF) was determined using a low-angle laser light scattering combined with a differential refractometer and a uv detector. The protein samples were applied to these detectors through a gel filtration column by a high-performance liquid chromatographic pump. The Chinese hamster ovary (CHO) cell-derived SCF gave a molecular weight of 53,000 for the entire molecule and 35,000 for the protein moiety only at pH 7.0, indicating that the CHO cell-derived protein is glycosylated by 34%. Since the molecular weight of the polypeptide is 18,600, the results demonstrate that the CHO cell-derived SCF forms a dimer. The molecular weight of Escherichia coli-derived SCF was determined to be 39,000, similar to the above value (35,000). Essentially identical molecular weights were obtained at pH 3.0, indicating no dissociation of the dimer.
Arakawa
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Amgen Inc., Thousand Oaks, California 91320.
Langley
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Kameyama
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Takagi
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D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D006031:Glycosylation; D016298:Hematopoietic Cell Growth Factors / Q000737:chemistry*; D006801:Humans; D007834:Lasers; D008027:Light; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000737:chemistry; D012542:Scattering, Radiation; D019089:Stem Cell Factor
D016298:Hematopoietic Cell Growth Factors; D011994:Recombinant Proteins; D019089:Stem Cell Factor