Cloning of mouse prostaglandin transporter PGT cDNA: Species-specific substrate affinities

Michael L. Pucci, Yi Bao, Brenda Chan, Shigekazu Itoh, Run Lu, Neal G. Copeland, Debra J. Gilbert, Nancy A. Jenkins, Victor L. Schuster

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Abstract

We recently identified and/or cloned the PG transporter PGT in the rat (rPGT) (Kanai, N., R. Lu, J. A. Satriano, Y. Bao, A. W. Wolkoff, and V. L. Schuster, Science 268: 866-869, 1995) and the human (hPGT) (Lu, R., and V. L. Schuster, J. Clin. Invest. 98: 1142-1149, 1996). Here we have cloned and expressed the mouse PGT (mPGT) cDNA. The tissue distribution of mPGT mRNA expression is significantly more restricted than that of rPGT and hPGT mRNA. Although the deduced amino acid sequence of mPGT is similar to the rat (91% identity) and human (82% identity) homologues, it has three regions of dissimilarity: amino acids 128-163 and 283-298, and valine 610 and isoleucine 611 (predicted to lie within putative transmembrane span 12). Affinities of hPGT, rPGT, and mPGT for several PG substrates differed, with hPGT having the highest [low Michaelis constant (K(m))] and mPGT the lowest affinity. A chimeric protein, linking the N-terminal domain of mPGT with the C-terminal domain of hPGT, had affinity for PGE2 indistinguishable from that of hPGT, indicating that the C-terminal domain dictates K(m). We mutagenized mouse valine 610 and isoleucine 611 to their corresponding human residues (methionine and glycine, respectively); however, these changes did not convert the inhibition constant of mPGT to that of hPGT. The mouse gene was localized to chromosome 9 in a region syntenic with the region of human chromosome 3 containing the hPGT gene. These studies highlight the species- dependence of tissue expression and function of PGT and lay the groundwork for the use of the mouse as a model system for the study of PGT function.

Original languageEnglish (US)
Pages (from-to)R734-R741
JournalAmerican Journal of Physiology - Regulatory Integrative and Comparative Physiology
Volume277
Issue number3 46-3
StatePublished - Sep 1 1999

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Keywords

  • Biological transport
  • Carrier proteins
  • Interspecific mouse backcross mapping
  • Molecular cloning

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)

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