Characterization and quantification of pyridoxal 5'-phosphate-extracted nuclear progesterone receptor

Tong J. Chen, Richard G. MacDonald, William F. Robidoux, Wendell W. Leavitt

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Pyridoxal phosphate extracts progesterone receptors from the nuclear fraction of hamster uterus. Optimal extraction occurs after 2 h at 2 C with 5 mM pyridoxal phosphate. At millimolar concentrations, pyridoxal phosphate also extracts non-receptor proteins from the nuclei, in amounts equivalent to those released by high salt (0.5 M KO) treatment (~10 mg protein per gram tissue). Thus, although pyridoxal phosphate is a useful probe for studying the interaction of steroid receptors with nuclear components, it does not selectively extract nuclear steroid receptors. The pyridoxal phosphate-extracted nuclear progesterone receptor had a sedimentation coefficient of 3.3S. Upon reduction with sodium borohydride, this species was converted to a 2.5S molecule. Sodium borohydride reduction of the pyridoxal phosphate-extracted nuclear receptor also caused a significant change in the steroid binding specificity of the molecule. The steroid specificity of the unreduced pyridoxal phosphate-extracted nuclear receptor was virtually identical to that of the KCl-extracted nuclear receptor. In contrast, deoxycorticosterone and 5α-pregnanedione competed 3- to 8-fold less effectively with progesterone for binding to the reduced form of pyridoxal phosphate-extracted nuclear receptor as compared to the other receptor forms.

Original languageEnglish (US)
Pages (from-to)1023-1028
Number of pages6
JournalJournal of Steroid Biochemistry
Volume14
Issue number10
DOIs
StatePublished - Oct 1981

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Pyridoxal Phosphate
Progesterone Receptors
Cytoplasmic and Nuclear Receptors
Steroid Receptors
Pregnanediones
Steroids
Desoxycorticosterone
Molecules
Sedimentation
Cricetinae
Uterus
Progesterone
Proteins
Salts
Tissue

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology

Cite this

Characterization and quantification of pyridoxal 5'-phosphate-extracted nuclear progesterone receptor. / Chen, Tong J.; MacDonald, Richard G.; Robidoux, William F.; Leavitt, Wendell W.

In: Journal of Steroid Biochemistry, Vol. 14, No. 10, 10.1981, p. 1023-1028.

Research output: Contribution to journalArticle

Chen, Tong J. ; MacDonald, Richard G. ; Robidoux, William F. ; Leavitt, Wendell W. / Characterization and quantification of pyridoxal 5'-phosphate-extracted nuclear progesterone receptor. In: Journal of Steroid Biochemistry. 1981 ; Vol. 14, No. 10. pp. 1023-1028.
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