Purification and characterization of 17α,20β-dihydroxy-4-pregnen-3-one binding protein from plasma of rainbow trout, Oncorhynchus mykiss

Michiyasu Yoshikuni, Hisami Matsushita, Naoki Shibata, Yoshitaka Nagahama

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

A 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-DP) (the natural maturation-inducing hormone of salmonid fish) binding protein (DBP) was purified from rainbow trout plasma. It had an apparent molecular weight of 110 kDa on native PAGE and was composed of two subunits with molecular weights of 50 and 55 kDa on SDS-PAGE. Enzymatic digestion of sugar chains converted both subunits to a single peptide with a molecular weight of 42.5 kDa. Scatchard analysis of 17α,20β-DP binding to purified DBP showed the presence of a single binding site with a K(d) of 21 nM and B(max) of 5 nmol/mg of protein. The affinities of various steroids were estimated by the displacement of [3H]17α,20β-DP binding in the decreasing order of 17α,20β-DP, testosterone, 17α-hydroxyprogesterone, progesterone, estradiol-17β, 17α,20β,21-trihydroxy-4-pregnen-3-one, and cortisol. We conclude that DBP is a sex-hormone binding globulin in rainbow trout.

Original languageEnglish
Pages (from-to)189-196
Number of pages8
JournalGeneral and Comparative Endocrinology
Volume96
Issue number2
DOIs
Publication statusPublished - Nov 1994

Fingerprint

Oncorhynchus mykiss
binding proteins
Carrier Proteins
Molecular Weight
molecular weight
17-hydroxyprogesterone
Fish Proteins
17-alpha-Hydroxyprogesterone
Native Polyacrylamide Gel Electrophoresis
Sex Hormone-Binding Globulin
testosterone
steroids
cortisol
Progesterone
Hydrocortisone
Testosterone
estradiol
Polyacrylamide Gel Electrophoresis
binding sites
Digestion

All Science Journal Classification (ASJC) codes

  • Animal Science and Zoology
  • Endocrinology

Cite this

Purification and characterization of 17α,20β-dihydroxy-4-pregnen-3-one binding protein from plasma of rainbow trout, Oncorhynchus mykiss. / Yoshikuni, Michiyasu; Matsushita, Hisami; Shibata, Naoki; Nagahama, Yoshitaka.

In: General and Comparative Endocrinology, Vol. 96, No. 2, 11.1994, p. 189-196.

Research output: Contribution to journalArticle

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abstract = "A 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-DP) (the natural maturation-inducing hormone of salmonid fish) binding protein (DBP) was purified from rainbow trout plasma. It had an apparent molecular weight of 110 kDa on native PAGE and was composed of two subunits with molecular weights of 50 and 55 kDa on SDS-PAGE. Enzymatic digestion of sugar chains converted both subunits to a single peptide with a molecular weight of 42.5 kDa. Scatchard analysis of 17α,20β-DP binding to purified DBP showed the presence of a single binding site with a K(d) of 21 nM and B(max) of 5 nmol/mg of protein. The affinities of various steroids were estimated by the displacement of [3H]17α,20β-DP binding in the decreasing order of 17α,20β-DP, testosterone, 17α-hydroxyprogesterone, progesterone, estradiol-17β, 17α,20β,21-trihydroxy-4-pregnen-3-one, and cortisol. We conclude that DBP is a sex-hormone binding globulin in rainbow trout.",
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