Stabilization of the structure of horse plasma vitamin D binding protein by disulfide bonds.

R. C. Robinson, L. D. Burtnick

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Vitamin D binding protein (DBP) was isolated from horse plasma in a four-step procedure that involved Affi-Gel Blue affinity chromatography, gel filtration, hydroxylapatite chromatography, and anion exchange high-pressure liquid chromatography. The yield of DBP from 80 mL of plasma was 6-7 mg. Horse plasma DBP closely resembles other plasma DBPs, being a tryptophan-free protein of Mr 53,000. It is able to bind to and block the polymerization of monomeric actin. The secondary structure of DBP was calculated from circular dichroism measurements to be 39% alpha-helix, 42% beta-sheet, and 19% random coil. Circular dichroism and fluorescence studies revealed that the disulfide bonds of DBP contribute substantial structural stabilization to the molecule with respect to thermal denaturation. The thermal stability of DBP can be used to advantage. Incorporation of a brief treatment at 70 degrees C into the preparative scheme enables omission of one chromatographic step, without detectable alteration of the purified product.

Original languageEnglish
Pages (from-to)10-15
Number of pages6
JournalBiochemistry and cell biology = Biochimie et biologie cellulaire
Volume70
Issue number1
DOIs
Publication statusPublished - Jan 1992
Externally publishedYes

Fingerprint

Vitamin D-Binding Protein
Disulfides
Horses
Blood Proteins
Carrier Proteins
Stabilization
Plasmas
Cibacron Blue F 3GA
Circular Dichroism
Hot Temperature
High pressure liquid chromatography
Affinity chromatography
Denaturation
Durapatite
Chromatography
Affinity Chromatography
Tryptophan
Polymerization
Gel Chromatography
Anions

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

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