Identification of a candidate membrane protein for the basolateral peptide transporter of rat small intestine.
A candidate protein for the basolateral peptide transporter of rat jejunum is described. Vascular perfusion of the photoaffinity label, [4-azido-D-phe]-L-ala (2.5mM), had no effect on the transepithelial transport of the non-hydrolysable dipeptide D-phe-L-gln (1mM) from the lumen, its mucosal accumulation or wash-out into the vascular perfusate. When the label was perfused luminally, the transepithelial transport of D-phe-L-gln was inhibited by 38% (P<0.001) and accumulation increased by 62% (P<0.05). These data are consistent with those of a basolateral transporter that is strongly asymmetric in its substrate binding and transport properties. Labelling of basolateral membrane vesicles with [4-azido-3,5-3H-D-phe]-L-ala revealed that the majority of label was incorporated into a single protein of M(r)112+/-2 kDa and pI 6.5. MALDI-TOF analysis of tryptic digests of the protein followed by database searches established that this protein was novel with no obvious similarity to PepT1, the apical membrane transporter.
Department of Biology, University of York, P.O. Box 373, York YO10 5YW, UK. ejs9@york.ac.uk
E J
EJ
Shepherd
N
N
Lister
J A
JA
Affleck
J R
JR
Bronk
G L
GL
Kellett
I D
ID
Collier
P D
PD
Bailey
C A R
CA
Boyd
D000818:Animals; D001386:Azides / Q000494:pharmacology; D001692:Biological Transport; D002352:Carrier Proteins / Q000737:chemistry* / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism*; D019992:Databases as Topic; D004847:Epithelial Cells / Q000378:metabolism*; D020224:Expressed Sequence Tags; D007583:Jejunum / Q000378:metabolism; D008027:Light; D008297:Male; D013058:Mass Spectrometry; D008956:Models, Chemical; D000074011:Peptide Transporter 1; D010477:Perfusion; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D019920:Photoaffinity Labels / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D027981:Symporters; D013997:Time Factors; D014357:Trypsin / Q000494:pharmacology
D001386:Azides; D002352:Carrier Proteins; D000074011:Peptide Transporter 1; D019920:Photoaffinity Labels; C489940:Slc15a1 protein, rat; D027981:Symporters; C027162:4-azidophenylalanine; D010649:Phenylalanine; D014357:Trypsin