Specific binding of [3H]17α,20β-dihydroxy-4-pregnen-3-one to oocyte cortices of rainbow trout (Oncorhynchus mykiss)

Michiyasu Yoshikuni, Naoki Shibata, Yoshitaka Nagahama

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

Specific binding of [3H]17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-DP) to plasma membranes prepared from defolliculated oocytes of rainbow trout (Onchorhynchus mykiss) was identified and characterized. Binding was rapid and reached equilibrium in 30 min. 17α,20β-DP strongly inhibited [3H] 17α,20β-DP binding in a competitive manner. Scatchard analysis revealed two different binding sites: a high affinity binding site with a Kd of 18 nM and a Bmax of 0.2 pmoles/mg protein; and a low affinity binding site with a Kd of 0.5 μM and a Bmax of 1 pmoles/mg protein. This binding activity was successfully solubilized with n-heptyl-β-D-thioglucoside. [3H]17α,20β-DP binding to solubilized preparations reached equilibrium in 1h, and was competitively inhibited with 17α,20β-DP and 17α,20β,21-trihydroxy-4-pregnen-3-one. However, Scatchard analysis showed a single binding site with a Kd of 0.3 μM. The reason for the disappearance of the high affinity binding site in solubilized preparations remains unclear. These results demonstrate that a specific binding site for 17α,20β-DP exists in the plasma membrane of rainbow trout oocytes.

Original languageEnglish
Pages (from-to)15-24
Number of pages10
JournalFish Physiology and Biochemistry
Volume11
Issue number1-6
DOIs
Publication statusPublished - Jul 1 1993
Externally publishedYes

Fingerprint

Oncorhynchus mykiss
rainbow
Oocytes
binding sites
cortex
oocytes
Binding Sites
membrane
plasma
protein
Cell membranes
Thioglucosides
plasma membrane
Cell Membrane
site preparation
17,20-dihydroxy-4-pregnen-3-one
analysis
Proteins
proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Physiology
  • Aquatic Science

Cite this

Specific binding of [3H]17α,20β-dihydroxy-4-pregnen-3-one to oocyte cortices of rainbow trout (Oncorhynchus mykiss). / Yoshikuni, Michiyasu; Shibata, Naoki; Nagahama, Yoshitaka.

In: Fish Physiology and Biochemistry, Vol. 11, No. 1-6, 01.07.1993, p. 15-24.

Research output: Contribution to journalArticle

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abstract = "Specific binding of [3H]17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-DP) to plasma membranes prepared from defolliculated oocytes of rainbow trout (Onchorhynchus mykiss) was identified and characterized. Binding was rapid and reached equilibrium in 30 min. 17α,20β-DP strongly inhibited [3H] 17α,20β-DP binding in a competitive manner. Scatchard analysis revealed two different binding sites: a high affinity binding site with a Kd of 18 nM and a Bmax of 0.2 pmoles/mg protein; and a low affinity binding site with a Kd of 0.5 μM and a Bmax of 1 pmoles/mg protein. This binding activity was successfully solubilized with n-heptyl-β-D-thioglucoside. [3H]17α,20β-DP binding to solubilized preparations reached equilibrium in 1h, and was competitively inhibited with 17α,20β-DP and 17α,20β,21-trihydroxy-4-pregnen-3-one. However, Scatchard analysis showed a single binding site with a Kd of 0.3 μM. The reason for the disappearance of the high affinity binding site in solubilized preparations remains unclear. These results demonstrate that a specific binding site for 17α,20β-DP exists in the plasma membrane of rainbow trout oocytes.",
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