Isolation of vitamin B12-binding proteins using affinity chromatography. II. Purification and properties of a human granulocyte vitamine B12-binding protein.
نویسندگان
چکیده
A vitamin Blz-binding protein has been isolated from human granulocytes derived from patients with chronic granulocytic leukemia. Utilizing affinity chromatography as the sole purification technique, the protein was purified 9,860-fold with a yield of over 90% and was homogeneous based on polyacrylamide disc gel electrophoresis, sedimentation equilibrium ultracentrifugation, and sodium dodecyl sulfate polyacrylamide gel electrophoresis. The protein binds 34.9 pg of vitamin BIG per mg of protein and has a single vitamin Blz-binding site. The molecular weight determined by sedimentation equilibrium ultracentrifugation was 56,000, whereas that determined by amino acid and carbohydrate analysis was 58,200. The protein contains 33 % carbohydrate which accounts for the elevated molecular weight values (121,000 to 138,000) obtained using gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis.
منابع مشابه
Isolation of vitamin B12-binding proteins using affinity chromatography. I. Preparation and properties of vitamin B12-sepharose.
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ورودعنوان ژورنال:
- The Journal of biological chemistry
دوره 247 23 شماره
صفحات -
تاریخ انتشار 1972