Involvement of SULT1A3 in elevated sulfation of 4-hydroxypropranolol in Hep G2 cells pretreated with β-naphthoflavone

Junko Miyano, Shigeo Yamamoto, Nobumitsu Hanioka, Shizuo Narimatsu, Tsutomu Ishikawa, Kenichiro Ogura, Tadashi Watabe, Masuhiro Nishimura, Nobuhiko Ueda, Shinsaku Naito

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Abstract

Pretreatment of Hep G2 cells with β-naphthoflavone (BNF 1-25 μM) significantly increased cytosolic sulfation activities of 4-hydroxypropranolol (4-OH-PL) racemate. The profile was similar to those of sulfations towards dopamine and triiodothyronine in the same cytosolic fractions. Kinetic studies of 4-OH-PL sulfation in Hep G2 cytosolic fractions revealed that Vmax values increased but apparent Km values remained unchanged following the BNF pretreatment. Among five recombinant human SULT isoforms (SULT1A1, -1A3, -1B1, -1E1 and -2A1) examined, only SULT2A1 did not show 4-OH-PL sulfation activities under the conditions used. SULT1A3 and -1E1 exhibited an enantioselectivity of 4-OH-R-PL sulfation > 4-OH-S-PL sulfation, which agreed with that of BNF-pretreated Hep G2 cells as well as of nontreated cells, whereas SULT1A1 and -1B1 showed a reversed enantioselectivity (R <S). In kinetic studies of 4-OH-PL sulfations by four kinds of human SULT isoforms, apparent Km values for SULT1A3 were the lowest, and the parameters were close to those of Hep G2 cytosolic fractions. Real time RT-PCR using TaqMan probes demonstrated that the mRNA levels of SULT1A3 increased following BNF pretreatment, which paralleled the results from Western blotting showing the elevated levels of SULT1A3 proteins. These results suggest that the induction of SULT1A3 is mainly responsible for the elevated 4-OH-PL sulfation activities following the pretreatment of Hep G2 cells with BNF.

Original languageEnglish
Pages (from-to)941-950
Number of pages10
JournalBiochemical Pharmacology
Volume69
Issue number6
DOIs
Publication statusPublished - Mar 15 2005

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Hep G2 Cells
Enantioselectivity
Protein Isoforms
Kinetics
Triiodothyronine
monoamine-sulfating phenol sulfotransferase
4-hydroxypropranolol
hydroxide ion
Real-Time Polymerase Chain Reaction
Dopamine
Western Blotting
Messenger RNA
Proteins

Keywords

  • β-Naphthoflavone
  • 4-Hydroxypropranolol
  • Hep G2 cell line
  • SULT1A3

ASJC Scopus subject areas

  • Pharmacology

Cite this

Involvement of SULT1A3 in elevated sulfation of 4-hydroxypropranolol in Hep G2 cells pretreated with β-naphthoflavone. / Miyano, Junko; Yamamoto, Shigeo; Hanioka, Nobumitsu; Narimatsu, Shizuo; Ishikawa, Tsutomu; Ogura, Kenichiro; Watabe, Tadashi; Nishimura, Masuhiro; Ueda, Nobuhiko; Naito, Shinsaku.

In: Biochemical Pharmacology, Vol. 69, No. 6, 15.03.2005, p. 941-950.

Research output: Contribution to journalArticle

Miyano, J, Yamamoto, S, Hanioka, N, Narimatsu, S, Ishikawa, T, Ogura, K, Watabe, T, Nishimura, M, Ueda, N & Naito, S 2005, 'Involvement of SULT1A3 in elevated sulfation of 4-hydroxypropranolol in Hep G2 cells pretreated with β-naphthoflavone', Biochemical Pharmacology, vol. 69, no. 6, pp. 941-950. https://doi.org/10.1016/j.bcp.2004.12.012
Miyano, Junko ; Yamamoto, Shigeo ; Hanioka, Nobumitsu ; Narimatsu, Shizuo ; Ishikawa, Tsutomu ; Ogura, Kenichiro ; Watabe, Tadashi ; Nishimura, Masuhiro ; Ueda, Nobuhiko ; Naito, Shinsaku. / Involvement of SULT1A3 in elevated sulfation of 4-hydroxypropranolol in Hep G2 cells pretreated with β-naphthoflavone. In: Biochemical Pharmacology. 2005 ; Vol. 69, No. 6. pp. 941-950.
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abstract = "Pretreatment of Hep G2 cells with β-naphthoflavone (BNF 1-25 μM) significantly increased cytosolic sulfation activities of 4-hydroxypropranolol (4-OH-PL) racemate. The profile was similar to those of sulfations towards dopamine and triiodothyronine in the same cytosolic fractions. Kinetic studies of 4-OH-PL sulfation in Hep G2 cytosolic fractions revealed that Vmax values increased but apparent Km values remained unchanged following the BNF pretreatment. Among five recombinant human SULT isoforms (SULT1A1, -1A3, -1B1, -1E1 and -2A1) examined, only SULT2A1 did not show 4-OH-PL sulfation activities under the conditions used. SULT1A3 and -1E1 exhibited an enantioselectivity of 4-OH-R-PL sulfation > 4-OH-S-PL sulfation, which agreed with that of BNF-pretreated Hep G2 cells as well as of nontreated cells, whereas SULT1A1 and -1B1 showed a reversed enantioselectivity (R <S). In kinetic studies of 4-OH-PL sulfations by four kinds of human SULT isoforms, apparent Km values for SULT1A3 were the lowest, and the parameters were close to those of Hep G2 cytosolic fractions. Real time RT-PCR using TaqMan probes demonstrated that the mRNA levels of SULT1A3 increased following BNF pretreatment, which paralleled the results from Western blotting showing the elevated levels of SULT1A3 proteins. These results suggest that the induction of SULT1A3 is mainly responsible for the elevated 4-OH-PL sulfation activities following the pretreatment of Hep G2 cells with BNF.",
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AU - Hanioka, Nobumitsu

AU - Narimatsu, Shizuo

AU - Ishikawa, Tsutomu

AU - Ogura, Kenichiro

AU - Watabe, Tadashi

AU - Nishimura, Masuhiro

AU - Ueda, Nobuhiko

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AB - Pretreatment of Hep G2 cells with β-naphthoflavone (BNF 1-25 μM) significantly increased cytosolic sulfation activities of 4-hydroxypropranolol (4-OH-PL) racemate. The profile was similar to those of sulfations towards dopamine and triiodothyronine in the same cytosolic fractions. Kinetic studies of 4-OH-PL sulfation in Hep G2 cytosolic fractions revealed that Vmax values increased but apparent Km values remained unchanged following the BNF pretreatment. Among five recombinant human SULT isoforms (SULT1A1, -1A3, -1B1, -1E1 and -2A1) examined, only SULT2A1 did not show 4-OH-PL sulfation activities under the conditions used. SULT1A3 and -1E1 exhibited an enantioselectivity of 4-OH-R-PL sulfation > 4-OH-S-PL sulfation, which agreed with that of BNF-pretreated Hep G2 cells as well as of nontreated cells, whereas SULT1A1 and -1B1 showed a reversed enantioselectivity (R <S). In kinetic studies of 4-OH-PL sulfations by four kinds of human SULT isoforms, apparent Km values for SULT1A3 were the lowest, and the parameters were close to those of Hep G2 cytosolic fractions. Real time RT-PCR using TaqMan probes demonstrated that the mRNA levels of SULT1A3 increased following BNF pretreatment, which paralleled the results from Western blotting showing the elevated levels of SULT1A3 proteins. These results suggest that the induction of SULT1A3 is mainly responsible for the elevated 4-OH-PL sulfation activities following the pretreatment of Hep G2 cells with BNF.

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