Site-directed mutagenesis of the conserved tyrosine 151 of human placental NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase yields a catalytically inactive enzyme

C M Ensor; H H Tai

doi:10.1016/s0006-291x(05)80262-1

Site-directed mutagenesis of the conserved tyrosine 151 of human placental NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase yields a catalytically inactive enzyme

Biochem Biophys Res Commun. 1991 Apr 30;176(2):840-5. doi: 10.1016/s0006-291x(05)80262-1.

Authors

C M Ensor¹, H H Tai

Affiliation

¹ Division of Medicinal Chemistry and Pharmaceutics, College of Pharmacy, University of Kentucky, Lexington 40536-0082.

PMID: 2025296
DOI: 10.1016/s0006-291x(05)80262-1

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alanine / genetics
Amino Acid Sequence
Codon
Enzyme Activation
Escherichia coli / genetics
Gene Expression
Humans
Hydroxyprostaglandin Dehydrogenases / genetics*
Molecular Sequence Data
Mutagenesis, Site-Directed
Pregnancy Proteins / genetics
Tyrosine / genetics*

Substances

Codon
Pregnancy Proteins
Tyrosine
Hydroxyprostaglandin Dehydrogenases
15-hydroxyprostaglandin dehydrogenase
Alanine