Identification of a candidate membrane protein for the basolateral peptide transporter of rat small intestine

E. J. Shepherd, N. Lister, J. A. Affleck, J. R. Bronk, G. L. Kellett, I. D. Collier, P. D. Bailey, C. A R Boyd

Research output: Contribution to journalArticle

Abstract

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 (2.5 mM), had no effect on the transepithelial transport of the non-hydrolysable dipeptide D-phe-L-gln (1 mM) 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 Mr 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. © 2002 Elsevier Science (USA). All rights reserved.

Original languageEnglish
Pages (from-to)918-922
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume296
Issue number4
DOIs
Publication statusPublished - 2002

Fingerprint

Small Intestine
Membrane Proteins
Blood Vessels
Photoaffinity Labels
Protein Databases
Proteins
Membrane Transport Proteins
Dipeptides
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Jejunum
Perfusion
Membranes
peptide permease

Keywords

  • Oligopeptide
  • PepT1
  • Transport
  • Vascular perfusion

Cite this

Shepherd, E. J., Lister, N., Affleck, J. A., Bronk, J. R., Kellett, G. L., Collier, I. D., ... Boyd, C. A. R. (2002). Identification of a candidate membrane protein for the basolateral peptide transporter of rat small intestine. Biochemical and Biophysical Research Communications, 296(4), 918-922. https://doi.org/10.1016/S0006-291X(02)02021-1
Shepherd, E. J. ; Lister, N. ; Affleck, J. A. ; Bronk, J. R. ; Kellett, G. L. ; Collier, I. D. ; Bailey, P. D. ; Boyd, C. A R. / Identification of a candidate membrane protein for the basolateral peptide transporter of rat small intestine. In: Biochemical and Biophysical Research Communications. 2002 ; Vol. 296, No. 4. pp. 918-922.
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Shepherd, EJ, Lister, N, Affleck, JA, Bronk, JR, Kellett, GL, Collier, ID, Bailey, PD & Boyd, CAR 2002, 'Identification of a candidate membrane protein for the basolateral peptide transporter of rat small intestine', Biochemical and Biophysical Research Communications, vol. 296, no. 4, pp. 918-922. https://doi.org/10.1016/S0006-291X(02)02021-1

Identification of a candidate membrane protein for the basolateral peptide transporter of rat small intestine. / Shepherd, E. J.; Lister, N.; Affleck, J. A.; Bronk, J. R.; Kellett, G. L.; Collier, I. D.; Bailey, P. D.; Boyd, C. A R.

In: Biochemical and Biophysical Research Communications, Vol. 296, No. 4, 2002, p. 918-922.

Research output: Contribution to journalArticle

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AU - Affleck, J. A.

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AU - Bailey, P. D.

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AB - 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 (2.5 mM), had no effect on the transepithelial transport of the non-hydrolysable dipeptide D-phe-L-gln (1 mM) 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 Mr 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. © 2002 Elsevier Science (USA). All rights reserved.

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