Differential recognition of beta -lactam antibiotics by intestinal and renal peptide transporters, PEPT 1 and PEPT 2.
This study was initiated to determine if there are differences in the recognition of beta -lactam antibiotics as substrates between intestinal and renal peptide transporters, PEPT 1 and PEPT 2. Reverse transcription-coupled polymerase chain reaction and/or Northern blot analysis have established that the human intestinal cell line Caco-2 expresses PEPT 1 but not PEPT 2, whereas the rat proximal tubule cell line SKPT expresses PEPT 2 but not PEPT 1. Detailed kinetic analysis has provided unequivocal evidence for participation of PEPT 2 in SKPT cells in the transport of the dipeptide glycylsarcosine and the aminocephalosporin cephalexin. The substrate recognition pattern of PEPT 1 and PEPT 2 was studied with cefadroxil (a cephalosporin) and cyclacillin (a penicillin) as model substrates for the peptide transporters constitutively expressed in Caco-2 cells (PEPT 1) and SKPT cells (PEPT 2). Cyclacillin was 9-fold more potent than cefadroxil in competing with glycylsacosine for uptake via PEPT 1. In contrast, cefadroxil was 13-fold more potent than cyclacillin in competing with the dipeptide for uptake via PEPT 2. The substrate recognition pattern of PEPT 1 and PEPT 2 was also investigated using cloned human peptide transporters functionally expressed in HeLa cells. Expression of PEPT 1 or PEPT 2 in HeLa cells was found to induce H(+)-coupled cephalexin uptake in these cells. As was the case with Caco-2 cells and SKPT cells, the uptake of glycylsarcosine induced in HeLa cells by PEPT 1 cDNA and PEPT 2 cDNA was inhibitable by cyclacillin and cefadroxil. Again, the PEPT 1 cDNA-induced dipeptide uptake was inhibited more potently by cyclacillin than by cefadroxil, and the PEPT 2 cDNA-induced dipeptide uptake was inhibited more potently by cefadroxil than by cyclacillin. It is concluded that there are marked differences between the intestinal and renal peptide transporters in the recognition of beta -lactam antibiotics as substrates.
Ganapathy
M E
ME
Department of Medicine, Medical College of Georgia, Augusta 30912, USA.
Brandsch
M
M
Prasad
P D
PD
Ganapathy
V
V
Leibach
F H
FH
D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002434:Cefadroxil / Q000378:metabolism; D002478:Cells, Cultured; D002506:Cephalexin / Q000378:metabolism; D002511:Cephalosporins / Q000378:metabolism; D003493:Cyclacillin / Q000378:metabolism; D004151:Dipeptides / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007421:Intestine, Small / Q000166:cytology / Q000378:metabolism*; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D010406:Penicillins / Q000378:metabolism; D000074011:Peptide Transporter 1; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D027981:Symporters; D014616:Vaccinia virus / Q000235:genetics
D000900:Anti-Bacterial Agents; D002352:Carrier Proteins; D002511:Cephalosporins; D004151:Dipeptides; D010406:Penicillins; D000074011:Peptide Transporter 1; D012333:RNA, Messenger; D011994:Recombinant Proteins; C082186:SLC15A1 protein, human; C489940:Slc15a1 protein, rat; D027981:Symporters; C093694:hydrogen-coupled oligopeptide transporter PepT2; D002434:Cefadroxil; C004194:glycylsarcosine; D003493:Cyclacillin; D002506:Cephalexin