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Abstract
Membrane-bound dipeptidase (MBD) participates in the degradation of glutathione by cleaving the cysteinyl-glycine bond of cystinyl bisglycine (oxidized cysteinyl-glycine) following removal of a gamma-glutamyl group by gamma-glutamyl transpeptidase (GGT). In the mouse, MBD RNA is most abundant in small intestine, kidney, and lung and is represented by four distinct RNA species. These are generated by transcription from two promoters located 6 kilobases apart in the 5' flanking region of the gene and by the use of two different poly(A) addition sites. Promoter I is used primarily in small intestine and kidney, whereas promoter II is most active in lung and kidney. We found a discordance in the expected co-expression of MBD and GGT; as expected, MBD and GGT are both expressed at high levels in the kidney and small intestine. However, in the lung, MBD is expressed at high levels, whereas GGT is almost undetectable. The reverse is true in the seminal vesicles and fetal liver. Thus, although both enzymes may function in concert to metabolize glutathione in kidney and small intestine, in other tissues they appear to act independently, suggesting that they have independent roles in other biological processes.
Highlights
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) U48387, U48388, U48389, and U48390
We have examined tissue-specific steady-state Membrane-bound dipeptidase (MBD) RNA levels on a comparative basis with group by ␥-glutamyl transpeptidase (GGT), identified four distinct MBD RNA species, and demonstrated that they are encoded by a single MBD gene
Comparison of MBD and GGT Expression—Because MBD and GGT are both ectoenzymes and function in concert to metabolize GSH, we evaluated the tissue distribution of steady state RNA levels of these two enzymes on a comparative basis (Fig. 1, A and B)
Summary
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) U48387, U48388, U48389, and U48390. In Situ Hybridization—Sequences representing 52 bp of the 5Ј common untranslated region and 95 bp of the coding region were amplified by RT-PCR using mouse kidney RNA as the template using primer 5 (see under “Construction of Mouse Kidney MBD cDNA Clones by RACE”) as the antisense primer and primer 7, 5Ј-ACGGAGGTGCCAAAGCCGCTG-3Ј, as the sense primer. Kidney and small intestine both expressed high levels of MBD and GGT.
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