20.BETA.-Hydroxysteroid dehydrogenase of neonatal pig testis. Localization in cytosol fraction and comparison with the enzyme from other species.

Nakajin, Shizuo; Ohno, Shuji; Takahashi, Masayuki; Nishimura, Kiyoshi; Shinoda, Masato

doi:10.1248/cpb.35.2490

Cited by 19 publications

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“…Moreover, studies using cell-free systems in amago salmon (our unpublished data) and Japanese eel (20) have demonstrated that the 20β-HSD activity in ovarian follicles is mainly located in the membrane-bound fractions of mitochondria and microsomes, with lower levels detected in the cytosolic fraction, which is inconsistent with the observation of CR/20β-HSD in the cytosol of pigs (2). Therefore, it seems premature to conclude that CR/20β-HSD is responsible for DHP production in the ovarian follicles of salmonids during FOM.…”

Section: Introductionmentioning

confidence: 60%

17β-HSD type 12-like is responsible for maturation-inducing hormone synthesis during oocyte maturation in masu salmon¹²

et al. 2016

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Abbreviated title: 17β-HSD12-like responsible for DHP production Key terms: 17α,20β-dihydroxy-4-pregnen-3-one, 20β-hydroxysteroid dehydrogenase, 17β-hydroxysteroid dehydrogenase, Oocyte maturation, Granulosa, Masu salmon, Oncorhynchus masou AbstractThe maturation-inducing hormone 17α,20β-dihydroxy-4-pregnen-3-one (DHP) was first identified in the amago salmon. However, although carbonyl reductase-like 20β-hydroxysteroid dehydrogenase (CR/20β-HSD) was reported to convert 17α-hydroxyprogesterone (17OHP) to DHP in rainbow trout, we previously found that CR/20β-HSD mRNA was not up-regulated in stimulated granulosa cells from masu salmon, which suggested that DHP is synthesized by a different enzyme. Accordingly, the present study aimed to identify the specific 20β-hydroxysteroid dehydrogenase (20β-HSD) responsible for DHP production by granulosa cells during final oocyte maturation in masu salmon. RNA-sequencing was performed on granulosa layers that were isolated from ovarian follicles at one month before ovulation and incubated with or without forskolin, which was used to mimic luteinizing hormone, and ~12 million reads were obtained, which yielded 71,062 contigs of > 100 bp. tBlastx analysis identified one contig (#f103496) as similar to 17β-hydroxysteroid dehydrogenase type 12 (hsd17β12); however, because the fulllength #f103496 sequence was different from hsd17β12, it was termed to hsd17β12-like (hsd17β12l). We found that mammalian cells transfected with full-length hsd17β12l exhibited considerable 20β-HSD activity, as indicated by efficient conversion of exogenous 17OHP to DHP. In addition, we found that hsd17β12l mRNA levels were consistently low in follicles during vitellogenic growth; however, the levels increased significantly during final oocyte maturation. The levels of hsd17β12l mRNA were also considerably increased in granulosa layers in which 20β-HSD activity was induced by salmon pituitary extract. Therefore, we suggest that hsd17β12l, not CR/20β-HSD, is the 20β-HSD responsible for DHP production by granulosa cells in masu salmon during final oocyte maturation.

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Section: Introductionmentioning

confidence: 60%

17β-HSD type 12-like is responsible for maturation-inducing hormone synthesis during oocyte maturation in masu salmon¹²

et al. 2016

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“…A member of the SDR superfamily, the multifunctional enzyme porcine testicular carbonyl reductase (PTCR)/3␣/␤, 20␤-hydroxysteroid dehydrogenase (20␤-HSD), catalyzes the NADPH-dependent reduction of ketones on androgens, progestins, and prostaglandins, as well as aldehydes and ketones on a large number of xenobiotics (2,3). The high level of 20␤-HSD activity in the enzyme is demonstrated by the reduction of 20-carbonyl groups of C 21 -steroids, such as conversion of 17␣-hydroxyprogesterone to 17␣,20␤-dihydroxy-4-pregnen-3-one, which is present in pig testes during the neonatal stage (4,5). Purified PTCR also shows vigorous 3␣-and 3␤-HSD activities with 5␣-androstan-17␤-ol-3-one (5␣-dihydrotestosterone) as a substrate (6).…”

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confidence: 99%

Porcine Carbonyl Reductase

Ghosh

Sawicki

Плетнев

et al. 2001

Journal of Biological Chemistry

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Porcine testicular carbonyl reductase (PTCR) belongs to the short chain dehydrogenases/reductases (SDR) superfamily and catalyzes the NADPH-dependent reduction of ketones on steroids and prostaglandins. The enzyme shares nearly 85% sequence identity with the NADPH-dependent human 15-hydroxyprostaglandin dehydrogenase/carbonyl reductase. The tertiary structure of the enzyme at 2.3 Å reveals a fold characteristic of the SDR superfamily that uses a Tyr-Lys-Ser triad as catalytic residues, but exhibits neither the functional homotetramer nor the homodimer that distinguish all SDRs. It is the first known monomeric structure in the SDR superfamily. In PTCR, which is also active as a monomer, a 41-residue insertion immediately before the catalytic Tyr describes an all-helix subdomain that packs against interfacial helices, eliminating the fourhelix bundle interface conserved in the superfamily. An additional anti-parallel strand in the PTCR structure also blocks the other strand-mediated interface. These novel structural features provide the basis for the scaffolding of one catalytic site within a single molecule of the enzyme.

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“…Gonadotropin influences DHP production by inducing the activity of the enzyme 20βhydroxysteroid dehydrogenase (20β-HSD) (Nagahama 1987b) to catalyze the reduction of 20carbonyl groups to 20β-hydroxylated products (Nagahama and Yamashita 2008). An enzyme with 20β-HSD activity was first isolated from the neonatal pig testis, showing strong 17αhydroxyprogesterone (17OHP) conversion activity to produce DHP (Nakajin et al, 1987), and its cDNA sequence shows high similarity to human carbonyl reductase (Tanaka et al, 1992), which is thought to catalyze the reduction of carbonyl compounds with NAD(P)H-dependent oxidoreductase activity (Tanaka et al, 2002). The significance of the identity between 20β-HSD and carbonyl reductase is unclear, since enzymes with similar functions catalyze the reduction of carbonyl groups to form corresponding products (Kazeto et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

17β-Hydroxysteroid dehydrogenase type 12 is responsible for maturation-inducing steroid synthesis during oocyte maturation in Nile tilapia

Aranyakanont

Ijiri

Hasegawa

et al. 2020

General and Comparative Endocrinology

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17α, 20β-Dihydroxy-4-pregnen-3-one (DHP) is a maturation-inducing steroid in many teleost fish. Carbonyl reductase-like 20β-hydroxysteroid dehydrogenase (CR/20β-HSD) is a candidate enzyme responsible for DHP production during oocyte maturation in various fish, including Nile tilapia. However, a novel type of 17β-hydroxysteroid dehydrogenase, type 12like (17β-HSD12L), is responsible for DHP production during oocyte maturation in masu salmon. 17β-HSD12 (presumably orthologous to salmon 17β-HSD12L) has been detected in Nile tilapia; however, its enzymatic activity and specific ability to convert the DHP substrate 17α-hydroxyprogesterone (17OHP) have not been examined. This study aimed to determine whether CR/20β-HSD or 17β-HSD12 is responsible for DHP production during oocyte maturation in the Nile tilapia. Mammalian expression vectors containing tilapia hsd17b12 or CR/20bhsd were transfected into HEK293T cells, followed by incubation with 17OHP.HEK293T cells transfected with hsd17b12 exhibited a strong ability to convert exogenous 17OHP to DHP (73.8% yield). Cells transfected with CR/20bhsd or the control vector converted only 7.4% and 7.5% of 17OHP to DHP, respectively. In addition, based on LC-MS/MS analyses, 17β-HSD12 did not convert any substrates other than 17OHP, including DHP, adrenosterone, androstenedione, estrone, testosterone, 11-ketotestosterone, and estradiol-17β. CR/20β-HSD showed strong 17β-HSD oxidoreductase activity especially with adrenosterone and androstenedione. Tissue-specific hsd17b12 expression analyzed by RT-PCR showed that hsd17b12 mRNA was strongest amplification in full-grown follicles.Finally, full-grown ovarian follicles were incubated with salmon pituitary extract (SPE, 100 µg/mL) or human chorionic gonadotropin (HCG,100 IU/mL) to induce 20β-HSD activity in vitro, and enzyme activity was assessed by co-incubation with 100 ng/mL 17OHP for 2, 4, 8, and 16 h. Conversion of 17OHP to DHP by ovarian follicles incubated with SPE and HCG peaked at 16 h, subsequent with increased follicular hsd17b12 mRNA levels, which were significantly higher than those in control incubations. However, the levels of CR/20bhsd mRNA remained low and did not differ among time points. The present study strongly suggests that 17β-HSD12, and not CR/20β-HSD, is the 20β-HSD responsible for DHP production by ovarian follicles during oocyte maturation in Nile tilapia.

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20.BETA.-Hydroxysteroid dehydrogenase of neonatal pig testis. Localization in cytosol fraction and comparison with the enzyme from other species.

Abstract: 20ƒÀ-Hydroxysteroid dehydrogenase (20ƒÀ-HSD) was found in neonatal pig testes. The enzyme

Cited by 19 publications

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17β-HSD type 12-like is responsible for maturation-inducing hormone synthesis during oocyte maturation in masu salmon¹²

17β-HSD type 12-like is responsible for maturation-inducing hormone synthesis during oocyte maturation in masu salmon¹²

Porcine Carbonyl Reductase

17β-Hydroxysteroid dehydrogenase type 12 is responsible for maturation-inducing steroid synthesis during oocyte maturation in Nile tilapia

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20.BETA.-Hydroxysteroid dehydrogenase of neonatal pig testis. Localization in cytosol fraction and comparison with the enzyme from other species.

Abstract: 20ƒÀ-Hydroxysteroid dehydrogenase (20ƒÀ-HSD) was found in neonatal pig testes. The enzyme

Cited by 19 publications

References 0 publications

17β-HSD type 12-like is responsible for maturation-inducing hormone synthesis during oocyte maturation in masu salmon12

17β-HSD type 12-like is responsible for maturation-inducing hormone synthesis during oocyte maturation in masu salmon12

Porcine Carbonyl Reductase

17β-Hydroxysteroid dehydrogenase type 12 is responsible for maturation-inducing steroid synthesis during oocyte maturation in Nile tilapia

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17β-HSD type 12-like is responsible for maturation-inducing hormone synthesis during oocyte maturation in masu salmon¹²

17β-HSD type 12-like is responsible for maturation-inducing hormone synthesis during oocyte maturation in masu salmon¹²