Site-directed mutagenesis of the conserved tyrosine 151 of human placental NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase yields a catalytically inactive enzyme.
NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH) is a key enzyme involved in the biological inactivation of the prostaglandins. The cDNA for human placental 15-PGDH has been expressed in Escherichia coli. Site-directed mutagenesis was used to convert a highly conserved tyrosine at position 151 in 15-PGDH to an alanine. The DNA coding for this alanine mutant 15-PGDH was expressed in E. coli. Western blot analysis indicated that this mutant protein was expressed in amounts comparable to the wild type enzyme in bacteria, however no 15-PGDH activity could be detected. This result indicates that tyrosine 151 in 15-PGDH is essential for activity.
Ensor
C M
CM
Division of Medicinal Chemistry and Pharmaceutics, College of Pharmacy, University of Kentucky, Lexington 40536-0082.
Tai
H H
HH
D000409:Alanine / Q000235:genetics; D000595:Amino Acid Sequence; D003062:Codon; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D006911:Hydroxyprostaglandin Dehydrogenases / Q000235:genetics*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011257:Pregnancy Proteins / Q000235:genetics; D014443:Tyrosine / Q000235:genetics*
D003062:Codon; D011257:Pregnancy Proteins; D014443:Tyrosine; D006911:Hydroxyprostaglandin Dehydrogenases; C035859:15-hydroxyprostaglandin dehydrogenase; D000409:Alanine