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Volume 270, Number 12, Issue of March 24, 1995 pp. 6456-6463
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Human Intestinal H/Peptide Cotransporter
CLONING, FUNCTIONAL EXPRESSION, AND CHROMOSOMAL LOCALIZATION (*)

(Received for publication, July 18, 1994; and in revised form, December 22, 1994)

Rong Liang (1),  You-Jun Fei (1),  Puttur D. Prasad (1),  Sammanda Ramamoorthy (1),  Hong Han (2),  Teresa L. Yang-Feng (2),  Matthias A. Hediger (3),  Vadivel Ganapathy (1),  Frederick H. Leibach (1)(§)

From the  (1)Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta, Georgia 30912-2100, the (2)Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06510, and the (3)Departments of Medicine and Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115

ABSTRACT

In mammalian small intestine, a H-coupled peptide transporter is responsible for the absorption of small peptides arising from digestion of dietary proteins. Recently a cDNA clone encoding a H/peptide cotransporter has been isolated from a rabbit intestinal cDNA library (Fei, Y. J., Kanai, Y., Nussberger, S., Ganapathy, V., Leibach, F. H., Romero, M. F., Singh, S. K., Boron, W. F., and Hediger, M. A.(1994) Nature 368, 563-566). Screening of a human intestinal cDNA library with a probe derived from the rabbit H/peptide cotransporter cDNA resulted in the identification of a cDNA which when expressed in HeLa cells or in Xenopus laevis oocytes induced H-dependent peptide transport activity. The predicted protein consists of 708 amino acids with 12 membrane-spanning domains and two putative sites for protein kinase C-dependent phosphorylation. The cDNA-induced transport process accepts dipeptides, tripeptides, and amino beta-lactam antibiotics but not free amino acids as substrates. The human H/peptide cotransporter exhibits a high degree of homology (81% identity and 92% similarity) to the rabbit H/peptide cotransporter. But surprisingly these transporters show only a weak homology to the H-coupled peptide transport proteins present in bacteria and yeast. Chromosomal assignment studies with somatic cell hybrid analysis and in situ hybridization have located the gene encoding the cloned human H/peptide cotransporter to chromosome 13 q33q34.

FOOTNOTES

*
This work was supported by the National Institutes of Health Grant DK 28389. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked ``advertisement'' in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank(TM)/EMBL Data Bank with accession number(s) U13173[GenBank].

§
To whom correspondence should be addressed: Dept. of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta, GA 30912-2100. Tel.: 706-721-7661; Fax: 706-721-6608.

(^1)
R. Liang, Y.-J. Fei, P. D. Prasad, S. Ramamoorthy, H. Han, T. L. Yang-Feng, M. A. Hediger, V. Ganapathy, and F. H. Leibach, unpublished data.

ACKNOWLEDGEMENTS

We thank J. C. Liu for nucleotide sequencing and Joyce Hobson for secretarial assistance.

©1995 by The American Society for Biochemistry and Molecular Biology, Inc.

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