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Partial purification and characterization of a folatebinding protein from human choroid plexus

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Abstract

A folate-binding protein (binder) from human choroid plexus was solubilized with Triton X-100 and partially purified in three steps: (1) affinity chromatography, (2) Sephadex G-200 column chromatography, and (3) polyacrylamide gel electrophoresis. When the partially purified binder was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the binding activity was located in the region of the gel with a molecular weight between 45,000 and 60,000. The specific activity of the binder after the three purification steps was 1.2 μg folic acid/mg protein, a 316-fold purification. Binding activity of the partially purified binder decreased below pH 6.0 and above pH 8.0 was unaffected by treatment with ribonuclease or deoxyribonuclease, but was abolished with trypsin, chymotrypsin, or protease (Streptomyces griesus). The binding of folic acid to the human binder was inhibited by folate > H4-folate > methyl-H4-folate ≃dihydrofolate ≃ pteroic acid ≫ methotrexate ≃ aminopterin.

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Authors and Affiliations

  1. Division of Clinical Pharmacology, Department of Medicine and Pharmacology and Department of Pathology, University of Iowa College of Medicine, 52242, Iowa City, Iowa

    Suleiman A. Suleiman, Reynold Spector & Pasquale Cancilla

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  1. Suleiman A. Suleiman
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  2. Reynold Spector
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  3. Pasquale Cancilla
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Suleiman, S.A., Spector, R. & Cancilla, P. Partial purification and characterization of a folatebinding protein from human choroid plexus. Neurochem Res 6, 333–341 (1981). https://doi.org/10.1007/BF00964048

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  • DOI: https://doi.org/10.1007/BF00964048

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