Immunolocalization and pharmacological relevance of oligopeptide transporter PepT1 in intestinal absorption of beta-lactam antibiotics.
A polyclonal antibody (anti-PepT1/C) was raised against the rabbit intestinal H(+)-coupled oligopeptide transporter, PepT1. Anti-PepT1/C detected 70-80-kDa protein in crude membranes obtained from rabbit duodenum, jejunum and ileum. PepT1 was localized in the brush-border of the absorptive epithelial cells by subcellular fractionation of membranes on a sucrose density gradient and by immunohistochemistry using light and electron microscopy. Transport activity for cephalosporins and dipeptide expressed in Xenopus laevis oocytes injected with total mRNA obtained from rabbit small intestine was eliminated completely by prehybridization of the mRNA with antisense oligonucleotide against the 5'-coding region of rabbit PepT1 cDNA.
Sai
Y
Y
Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Kanazawa University, Ishikawa, Japan.
Tamai
I
I
Sumikawa
H
H
Hayashi
K
K
Nakanishi
T
T
Amano
O
O
Numata
M
M
Iseki
S
S
Tsuji
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000493:pharmacokinetics*; D002352:Carrier Proteins / Q000378:metabolism*; D004386:Duodenum / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism; D007082:Ileum / Q000378:metabolism; D015151:Immunoblotting; D007408:Intestinal Absorption; D007421:Intestine, Small / Q000378:metabolism / Q000648:ultrastructure; D007583:Jejunum / Q000378:metabolism / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D008871:Microvilli / Q000378:metabolism; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense; D000074011:Peptide Transporter 1; D011817:Rabbits; D027981:Symporters; D014982:Xenopus laevis; D047090:beta-Lactams
D000900:Anti-Bacterial Agents; D002352:Carrier Proteins; D016376:Oligonucleotides, Antisense; D000074011:Peptide Transporter 1; D027981:Symporters; D047090:beta-Lactams