Identification of three hydroxysteroid sulfotransferase isoenzymes in the rat liver

Mie Takahashi, Hiroomi Tamura, Sayuri Kondo, Kie Kobayashi, Michio Matsui

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Isolation of several hydroxysteroid sulfotransferase (HS-ST) cDNAs from the rat liver cDNA library has demonstrated the possible expression of these HS-ST isoforms in rat liver. We devised a method to detect unequivocally ST- 20 and ST-40 mRNAs by a reverse transcription-polymerase chain reaction, using specific primers. ST-40 mRNA was expressed only in liver, but ST-20 mRNA was present predominantly in the liver and slightly in extrahepatic tissues. On chromatofocusing, expressed ST-20 or ST-40 enzymes were eluted at approx. pH 8.2 and 5.7-4.7 or at pH 6.4-5.4, respectively. Chromatofocusing of adult female rat liver cytosols resolved HS-ST isoenzymes in a broad range of pH, and ST fractions A, B, C, D and E were eluted at approx. pH 8.2, 7.6, 7.5-6.8, 6.2 and 6.1-5.5, respectively. After PAP-agarose affinity column chromatography and SDS-polyacrylamide gel electrophoresis (PAGE), their N- terminal amino acid sequences were determined. ST isoenzymes present in fractions B and E showed identical N-terminal amino acid sequences with those of ST-21 and ST-20, respectively, whereas the ST isoenzymes present in fractions C and D had the same N-terminal amino acid sequence as those of ST- 40 (and/or ST-41). The results demonstrated the presence of at least three HS-ST isoenzymes in adult female rat liver.

Original languageEnglish
Pages (from-to)10-15
Number of pages6
JournalBiological and Pharmaceutical Bulletin
Volume21
Issue number1
Publication statusPublished - 1998

Fingerprint

Isoenzymes
Liver
Amino Acid Sequence
Messenger RNA
Agarose Chromatography
Gene Library
Affinity Chromatography
Cytosol
Reverse Transcription
alcohol sulfotransferase
Polyacrylamide Gel Electrophoresis
Protein Isoforms
Complementary DNA
Polymerase Chain Reaction
Enzymes

Keywords

  • Hydroxysteroid sulfotransferase
  • Isoenzyme
  • N-terminal amino acid sequence
  • Rat liver

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Identification of three hydroxysteroid sulfotransferase isoenzymes in the rat liver. / Takahashi, Mie; Tamura, Hiroomi; Kondo, Sayuri; Kobayashi, Kie; Matsui, Michio.

In: Biological and Pharmaceutical Bulletin, Vol. 21, No. 1, 1998, p. 10-15.

Research output: Contribution to journalArticle

Takahashi, Mie ; Tamura, Hiroomi ; Kondo, Sayuri ; Kobayashi, Kie ; Matsui, Michio. / Identification of three hydroxysteroid sulfotransferase isoenzymes in the rat liver. In: Biological and Pharmaceutical Bulletin. 1998 ; Vol. 21, No. 1. pp. 10-15.
@article{e0f31c4b29734423ac04b6ed275aa9bf,
title = "Identification of three hydroxysteroid sulfotransferase isoenzymes in the rat liver",
abstract = "Isolation of several hydroxysteroid sulfotransferase (HS-ST) cDNAs from the rat liver cDNA library has demonstrated the possible expression of these HS-ST isoforms in rat liver. We devised a method to detect unequivocally ST- 20 and ST-40 mRNAs by a reverse transcription-polymerase chain reaction, using specific primers. ST-40 mRNA was expressed only in liver, but ST-20 mRNA was present predominantly in the liver and slightly in extrahepatic tissues. On chromatofocusing, expressed ST-20 or ST-40 enzymes were eluted at approx. pH 8.2 and 5.7-4.7 or at pH 6.4-5.4, respectively. Chromatofocusing of adult female rat liver cytosols resolved HS-ST isoenzymes in a broad range of pH, and ST fractions A, B, C, D and E were eluted at approx. pH 8.2, 7.6, 7.5-6.8, 6.2 and 6.1-5.5, respectively. After PAP-agarose affinity column chromatography and SDS-polyacrylamide gel electrophoresis (PAGE), their N- terminal amino acid sequences were determined. ST isoenzymes present in fractions B and E showed identical N-terminal amino acid sequences with those of ST-21 and ST-20, respectively, whereas the ST isoenzymes present in fractions C and D had the same N-terminal amino acid sequence as those of ST- 40 (and/or ST-41). The results demonstrated the presence of at least three HS-ST isoenzymes in adult female rat liver.",
keywords = "Hydroxysteroid sulfotransferase, Isoenzyme, N-terminal amino acid sequence, Rat liver",
author = "Mie Takahashi and Hiroomi Tamura and Sayuri Kondo and Kie Kobayashi and Michio Matsui",
year = "1998",
language = "English",
volume = "21",
pages = "10--15",
journal = "Biological and Pharmaceutical Bulletin",
issn = "0918-6158",
publisher = "Pharmaceutical Society of Japan",
number = "1",

}

TY - JOUR

T1 - Identification of three hydroxysteroid sulfotransferase isoenzymes in the rat liver

AU - Takahashi, Mie

AU - Tamura, Hiroomi

AU - Kondo, Sayuri

AU - Kobayashi, Kie

AU - Matsui, Michio

PY - 1998

Y1 - 1998

N2 - Isolation of several hydroxysteroid sulfotransferase (HS-ST) cDNAs from the rat liver cDNA library has demonstrated the possible expression of these HS-ST isoforms in rat liver. We devised a method to detect unequivocally ST- 20 and ST-40 mRNAs by a reverse transcription-polymerase chain reaction, using specific primers. ST-40 mRNA was expressed only in liver, but ST-20 mRNA was present predominantly in the liver and slightly in extrahepatic tissues. On chromatofocusing, expressed ST-20 or ST-40 enzymes were eluted at approx. pH 8.2 and 5.7-4.7 or at pH 6.4-5.4, respectively. Chromatofocusing of adult female rat liver cytosols resolved HS-ST isoenzymes in a broad range of pH, and ST fractions A, B, C, D and E were eluted at approx. pH 8.2, 7.6, 7.5-6.8, 6.2 and 6.1-5.5, respectively. After PAP-agarose affinity column chromatography and SDS-polyacrylamide gel electrophoresis (PAGE), their N- terminal amino acid sequences were determined. ST isoenzymes present in fractions B and E showed identical N-terminal amino acid sequences with those of ST-21 and ST-20, respectively, whereas the ST isoenzymes present in fractions C and D had the same N-terminal amino acid sequence as those of ST- 40 (and/or ST-41). The results demonstrated the presence of at least three HS-ST isoenzymes in adult female rat liver.

AB - Isolation of several hydroxysteroid sulfotransferase (HS-ST) cDNAs from the rat liver cDNA library has demonstrated the possible expression of these HS-ST isoforms in rat liver. We devised a method to detect unequivocally ST- 20 and ST-40 mRNAs by a reverse transcription-polymerase chain reaction, using specific primers. ST-40 mRNA was expressed only in liver, but ST-20 mRNA was present predominantly in the liver and slightly in extrahepatic tissues. On chromatofocusing, expressed ST-20 or ST-40 enzymes were eluted at approx. pH 8.2 and 5.7-4.7 or at pH 6.4-5.4, respectively. Chromatofocusing of adult female rat liver cytosols resolved HS-ST isoenzymes in a broad range of pH, and ST fractions A, B, C, D and E were eluted at approx. pH 8.2, 7.6, 7.5-6.8, 6.2 and 6.1-5.5, respectively. After PAP-agarose affinity column chromatography and SDS-polyacrylamide gel electrophoresis (PAGE), their N- terminal amino acid sequences were determined. ST isoenzymes present in fractions B and E showed identical N-terminal amino acid sequences with those of ST-21 and ST-20, respectively, whereas the ST isoenzymes present in fractions C and D had the same N-terminal amino acid sequence as those of ST- 40 (and/or ST-41). The results demonstrated the presence of at least three HS-ST isoenzymes in adult female rat liver.

KW - Hydroxysteroid sulfotransferase

KW - Isoenzyme

KW - N-terminal amino acid sequence

KW - Rat liver

UR - http://www.scopus.com/inward/record.url?scp=0031936474&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031936474&partnerID=8YFLogxK

M3 - Article

VL - 21

SP - 10

EP - 15

JO - Biological and Pharmaceutical Bulletin

JF - Biological and Pharmaceutical Bulletin

SN - 0918-6158

IS - 1

ER -