Structure and Function of Human Nad+-Linked 15-Hydroxyprostaglandin Dehydrogenase

  • Hsin-Hsiung Tai
  • Charles Mark Ensor
  • Zhou Huiping
  • Yan Fengxiang
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 507)


Prostaglandins are rapidly metabolized in the body. The primary route of metabolism is initiated by NAD+-dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH) which catalyzes the reversible oxidation of 15(S)-hydroxyl group to a keto function resulting in a great loss of biological activities. 15-PGDH is therefore considered a key enzyme responsible for biological inactivation of prostaglandins. The enzyme is ubiquitously present in mammalian tissues. The first cDNA was cloned form human placenta and was found to have an open reading frame of 798 bp coding for 266 amino acids.’ The native enzyme is believed to be a dimer composed of identical subunits with a M.W. of 29 kDa. The enzyme utilizes a wide range of prostaglandins, lipoxins and hydroxy fatty acids as substrates and prefers NAD+ over NADP+ as a coenzyme.’ The enzyme is sensitive to sulfhydryl inhibitors indicating one or more cysteinyl residues are essential.’ The primary structure of 15-PGDH shows that it belongs to a superfamily of short chain dehydrogenases/reductases (SDR).3Sequence analysis indicates that an overall homology of 20% among SDRs with several conserved amino acid residues. Whether these conserved residues and cysteinyl residues are involved in catalytic activity and/or structural integrity remains to be determined. Most of these conserved residues reside in the first two thirds of the polypeptide chain suggesting that they might be involved in interacting with the common coenzyme NAD+. The C-terminal third shows little homology in sequences indicating that this segment appears to accommodate substrates of diversed structures.. This study is focused on using site-directed mutagenesis to examine the roles of several conserved and cysteinyl residues in catalytic activity and the involvement of C-terminal domain in enzyme and substrate interactions.


Mutant Enzyme Hydroxy Fatty Acid Conserve Amino Acid Residue Photoaffinity Label Inactive Enzyme 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    C.M. Ensor and H.H. Tai, Cloning and sequence analysis of the cDNA for human placental NAD*- dependent 15-hydroxyprostaglandin dehydrogenase, J. Biol. Chem. 265:14888 (1990)PubMedGoogle Scholar
  2. 2.
    D. Kung-Chao and H.H. Tai, NAD*-dependent 15-hydroxyprostaglandin dehydrogenase from porcine kidney, Biochim. Biophys. Acta 614:1 (1980)Google Scholar
  3. 3.
    H. Jornvall, B. Persson, M. Krook, S. Atrian, R. Gonzalez-Duarte, J. Jeffery and D. Ghosh, Short-chain dehydrogenases reductases (SDR). Biochemistry 34: 6003 (1995)PubMedCrossRefGoogle Scholar
  4. 4.
    R. Wierenga, P. Terpstra and W. Hol, Prediction of the occurrence of the ADP-binding 134-fold in proteins using an amino acid sequence fingerprint. J. Mol. Biol. 187:101 (1986)CrossRefGoogle Scholar
  5. 5.
    A. Chavan, C. Ensor, P. Wu, B. Haley and H.H. Tai, Photoaffinity labeling of human placental NAD’-linked 15-hydroxyprostaglandin dehydrogenase with [a-32P] 2N3NAD+.J. Biol. Chem. 268: 16437 (1993)PubMedGoogle Scholar
  6. 6.
    C. Ensor and H. H. Tai, Bacterial expression and site-directed mutagenesis of two critical residues (Tyrosine-151 and Lysine-155) of human placental NAD*-dependent 15-hydroxyprostaglandin dehydrogenase. Biochem. Biophys. Acta 1208: 151 (1994)PubMedGoogle Scholar
  7. 7.
    C. M. Ensor and H. H. Tai, Site-directed mutagenesis of the conserved serine 138 of human placental NAD+-dependent 15-hydroxyprostaglandin dehydrogenase to an alanine results in an inactive enzyme. Biochem. Biophys. Res. Commun. 220: 330 (1996)PubMedCrossRefGoogle Scholar
  8. 8.
    H. P. Zhou and H. H. Tai, Threonine 188 is critical for interaction with NAD* in Human NAD*-dependent 15-hydroxyprostaglandin dehydrogenase. Biochem. Biophys. Res. Commun. 257: 414 (1999)CrossRefGoogle Scholar
  9. 9.
    C.M. Ensor and H.H. Tai, Cysteine 182 is essential for enzymatic activity of human placental NAD* dependent 15-hydroxyprostaglandin dehydrogenase. Arch. Biochem. Biophys. 333: 117 (1996)PubMedCrossRefGoogle Scholar
  10. 10.
    N. Tanaka, T. Nonaka, T. Tanabe, T. Yoshimoto, D. Tsura and Y. Mitsui, Crystal structure of the binary and ternary complexes of 7a-hydroxysteroid dehydrogenase fromEscherichia Coli.Biochemistry 35: 7715(1996)Google Scholar

Copyright information

© Springer Science+Business Media New York 2002

Authors and Affiliations

  • Hsin-Hsiung Tai
    • 1
  • Charles Mark Ensor
    • 1
  • Zhou Huiping
    • 1
  • Yan Fengxiang
    • 1
  1. 1.Division of Pharmaceutical SciencesCollege of Pharmacy University of KentuckyLexington

Personalised recommendations