본문 바로가기
HOME> 논문 > 논문 검색상세

논문 상세정보

Role of Disulfide Bond of Arylsulfate Sulfotransferase in the Catalytic Activity

Kwon, Ae-Ran   (College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National UniversityUU0000691  ); Choi, Eung-Chil   (College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National UniversityUU0000691  );
  • 초록

    Bacterial arylsulfate sulfotransferase (ASST) catalyzes the transfer of sulfate group from a phenyl sulfate ester to a phenolic acceptor. The promoter region and the transcripti on start sites of Enterobacter amnigenus astA have been determined by primer extension analysis. Northern blot analysis resolved two mRNA species with lengths of 3.3 and 2.0 kb, which correspond to the distances between the transcriptional initiation sites and the two inverted repeat sequences (IRSs). By length, the 3.3 kb RNA could comprise the three-gene (astA with dsbA and dsbB) operon. ASST has three highly conserved cysteine residues. Reducing and non-reducing SDS-PAGE and activity staining showed that disulfide bond is needed for the activity of the enzyme. To identify the cysteine residues responsible for the disulfide bond formation, a series of Cys to Ser mutants has been constructed and the enzymatic activity was measured. Based on the results, we assumed that the first cysteine (Cys349) might be involved in disulfide bond mainly with the second cysteine (Cys445) and result in active conformation.


  • 주제어

    Arylsulfate sulfotransferase .   Enterobacter amnigenus .   Sodium dodecyl sulfate-polyacrylamide gel electrophoresis .   Disulfide bond .   Site-directed mutagenesis .   DsbA .   DsbB.  

  • 참고문헌 (28)

    1. Bardwell, J. C., McGovern, K., and Beckwith, J., Identification of a protein required for disulfide bond formation in vivo. Cell, 67, 581-589 (1991) 
    2. Feinberg, A. P. and Vogelstein, B., A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem., 132, 6-13 (1983) 
    3. Kahnert, A. and Kertesz M. A., Characterization of a sulfurregulated oxygenative alkylsulfatase from Pseudomonas putida S-313. J. Biol. Chem., 275, 31661-31667 (2000) 
    4. Kim, D. H., Kim, H. S., and Kobashi, K., Purification and characterization of novel sulfotransferase obtained from Klebsiella K-36, an intestinal bacterium of rat. J. Biochem., 112, 456-460 (1992) 
    5. Kim, D. H., Konishi, L., and Kobashi, K., Purification, characterization and reaction mechanism of novel arylsulfotransferase obtained from an anaerobic bacterium of human intestine. Biochem. Biophys. Acta, 872, 33-41 (1986) 
    6. Kobashi, K., Fukaya, Y., Kim, D. H., Akao, T., and Takebe, S., A novel type of arylsulfotransferase obtained from an anaerobic bacterium of human intestine. Arch. Biochem. Biophys., 245, 537-539 (1986) 
    7. Kunkel, T. A., Roberts J. D., and Zakour R. A., Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol., 154, 367-382 (1987) 
    8. Reeder, T. and Schleif, R., AraC protein can activate transcription from only one position and when pointed in only one direction. J. Mol. Biol., 231, 205-218 (1993) 
    9. Missiakas, D., C., Georgopoulos, and Raina S., Identification and characterization of the Escherichia coli gene dsbB, whose product is involved in the formation of disulfide bonds in vivo. Proc. Natl. Acad. Sci. U.S.A., 90, 7084-7088 (1993) 
    10. Kunkel, T. A., Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc. Natl. Acad. Sci. U.S.A., 82, 488-492 (1985) 
    11. Baek, M. C., Kim, S. K., Kim, D. H., Kim. B. K., and Choi, E. C., Cloning and sequencing of the Klebsiella K-36 astA gene, encoding an arylsulfate sulfotransferase. Microbiol. Immunol., 40, 531-537 (1996) 
    12. Dodgson, K. S. and Tudball, N., The metabolite of the sulfate group of potassium p-nitrophenyl[35S] sulfate. Biochem. J., 74, 154-159 (1960) 
    13. Sambrook, J., Fritsch, E. F., and Maniatis, T., Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. (1989) 
    14. Michael, H. M. and Bernard, L. H., Release of alkaline phosphatase from cells of Escherichia coli upon lysozyme spheroplast formation. J. Biol. Chem., 3, 1889-1893 (1964) 
    15. Sekura, R. D., Duffel, M. W., and Jakoby, W. B., Arylsulfotransferases. Methods Enzymol., 77, 197-206 (1981) 
    16. Laemmli, U. K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680- 685 (1970) 
    17. Kim, D. H. and Kobashi, K., The role of intestinal flora in metabolism of phenolic sulfate esters. Biochem. Pharmacol., 35, 3507-3510 (1986) 
    18. Kwon, A. R., Yun, H. J., and Choi, E. C., Kinetic mechanism and identification of the active site tyrosine residue in Enterobacter amnigenus arylsylfate sulfotrasferase. Biochem. Biophys. Res. Commun., 285, 526-529 (2001) 
    19. Harley, C. B. and Reynolds, R. P., Analysis of E. coli promoter sequences. Nucleic Acids Res., 15, 2343-2361 (1987) 
    20. Roy, A. B., Sulfotransferases. In Sulfation of Drugs and Related Compounds (Mulder, G. J., Ed.), pp. 131-185, CRC Press, Boca Raton, FL (1981) 
    21. Sanger, F., Nicklen, S., and Coulson, A. R., DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. U.S.A., 74, 5463-5467 (1977) 
    22. von Heijne, G. and Abrahmsen, L., Species-specific variation in signal peptide design. Implications for protein secretion in foreign hosts. FEABS Lett., 244, 439-446 (1989) 
    23. Neuhoff, V., Arold, N., Taube, D., and Ehrhardt, W., Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250. Electrophoresis, 9, 255-262 (1988) 
    24. Yao, R. and Guerry, P., Molecular cloning and site-specific mutagenesis of a gene involved in arylsulfate production in Campylobacter jejuni. J. Bacteriol., 178, 3335-3338 (1996) 
    25. Kang, J. W. Kwon, A. R., Kim, D. H., and Choi, E. C., Cloning and sequencing of the astA gene encoding arylsulfate sulfotransferase from Salmonella typhimurium. Bio. Pharm. Bull., 24, 570-574 (2001) 
    26. Lee, N. S., Kim, B. T., Kim, D. H., and Kobashi, K., Purification and reaction mechanism of arylsulfate sulfotransferase from Haemophilus K-12, a mouse intestinal bacterium. J. Biochem., 118, 796-801 (1995) 
    27. Chai, C. L. L. and Lowe, G., The mechanism and stereochemical course of sulfuryl transfer catalyzed by the acryl sulfotransferase from Eubacterium A-44. Bioorg. Chem., 20, 181-188 (1992) 
    28. Kwon, A. R., Oh, T. G., Kim, D. H., and Choi, E. C., Molecular cloning of the arylsulfate sulfotransferase gene and characterization of its product from Enterobacter amnigenus AR-37. Protein Expression Purif., 17, 366-372 (1999) 

 저자의 다른 논문

  • 권애란 (14)

    1. 1993 "리팜피신에 내성인 Streptococcus faecalis 균주의 개발" 약학회지 = Yakhak hoeji 37 (4): 350~355    
    2. 1993 "임상분리 Staphylococcus속 균주로부터 마크로라이드-린코사마이드-스트렙토그라민 B(MLS)계 항생물질에 대한 새로운 유도내성 유전자의 검색" The journal of applied pharmacology : the official journal of the Korean Society of Applied Pharmacology 1 (2): 177~182    
    3. 1994 "마크로라이드-린코사마이드-스트렙토그라민 B(MLS)계 항생물질에 대한 유도 내성" 약학회지 = Yakhak hoeji 38 (3): 293~299    
    4. 1997 "수소 생성 광합성 세균 Rhodobacter sphaeroides KS 56 분리" 한국식품영양학회지 = The Korean journal of food and nutrition 10 (4): 549~552    
    5. 1997 "광합성 세균 Rhodobacter sphaeroidea KS56에 의한 수소 생성" 東아시아食生活學會誌 = Journal of the East Asian Society of Dietary Life 7 (3): 325~329    
    6. 1998 "Distribution of Bacteria with the Arylsulfate Sulfotransferase Activity" Archives of pharmacal research : a publication of the Pharmaceutical Society of Korea 21 (4): 475~477    
    7. 1998 "한국형 유산균 Bifidobacterium 속 균주의 항생물질에 대한 감수성" 약학회지 = Yakhak hoeji 42 (6): 639~641    
    8. 1999 "리팜피신과 플로로퀴놀론계 항균제에 내성인 Bifidobacterium infantis OFR-525 균주" 약학회지 = Yakhak hoeji 43 (1): 124~127    
    9. 1999 "새로운 퀴놀론 항균제 DW-116의 살균 작용" 약학회지 = Yakhak hoeji 43 (1): 131~133    
    10. 2001 "Cloning, Sequence Analysis, and Characterization of the astA Gene Encoding an Arylsulfate Sulfotransferase from Citrobacter freundii" Archives of pharmacal research : a publication of the Pharmaceutical Society of Korea 24 (4): 316~322    
  • Choi, Eung-Chil (136)

 활용도 분석

  • 상세보기

    amChart 영역
  • 원문보기

    amChart 영역

원문보기

무료다운로드
유료다운로드

유료 다운로드의 경우 해당 사이트의 정책에 따라 신규 회원가입, 로그인, 유료 구매 등이 필요할 수 있습니다. 해당 사이트에서 발생하는 귀하의 모든 정보활동은 NDSL의 서비스 정책과 무관합니다.

원문복사신청을 하시면, 일부 해외 인쇄학술지의 경우 외국학술지지원센터(FRIC)에서
무료 원문복사 서비스를 제공합니다.

NDSL에서는 해당 원문을 복사서비스하고 있습니다. 위의 원문복사신청 또는 장바구니 담기를 통하여 원문복사서비스 이용이 가능합니다.

이 논문과 함께 출판된 논문 + 더보기