Functional characterization, localization, and molecular identification of rabbit intestinal N-amino acid transporter

Jamilur R. Talukder, Ramesh Kekuda, Prosenjit Saha, Puttur D Prasad, Vadivel Ganapathy, Uma Sundaram

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

We have characterized the Na-glutamine cotransporter in the rabbit intestinal crypt cell brush border membrane vesicles (BBMV). Substrate specificity experiments showed that crypt cell glutamine uptake is mediated by system N. Real-time PCR experiments showed that SN2 (SLC38A5) mRNA is more abundant in crypt cells compared with SN1 (SLC38A3), indicating that SN2 is the major glutamine transporter present in the apical membrane of the crypt cells. SN2 cDNA was obtained by screening a rabbit intestinal cDNA library with human SN1 used as probe. Rabbit SN2 cDNA encompassed a 473-amino-acid-long open reading frame. SN2 protein displayed 87% identity and 91% similarity to human SN2. Functional characterization studies of rabbit SN2 were performed by using vaccinia virusmediated transient expression system. Substrate specificity of the cloned transporter was identical to that of SN2 described in the literature and matched well with substrate specificity experiments performed using crypt cell BBMV. Cloned rabbit SN2, analogous to its human counterpart, is Li+ tolerant. Hill coefficient for Li+ activation of rabbit SN2-mediated uptake was 1. Taken together, functional data from the crypt cell BBMV and the cloned SN2 cDNA indicate that the crypt cell glutamine transport is most likely mediated by SN2.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume294
Issue number6
DOIs
StatePublished - Jun 1 2008

Fingerprint

Amino Acid Transport Systems
Rabbits
Glutamine
Microvilli
Substrate Specificity
Complementary DNA
Membranes
Vaccinia
Gene Library
Open Reading Frames
Real-Time Polymerase Chain Reaction
Cell Membrane
Amino Acids
Messenger RNA
Proteins

Keywords

  • Brush border membrane
  • Na glutamine cotransporter
  • Small intestine
  • System N-amino acid transporter

ASJC Scopus subject areas

  • Physiology
  • Hepatology
  • Gastroenterology
  • Physiology (medical)

Cite this

Functional characterization, localization, and molecular identification of rabbit intestinal N-amino acid transporter. / Talukder, Jamilur R.; Kekuda, Ramesh; Saha, Prosenjit; Prasad, Puttur D; Ganapathy, Vadivel; Sundaram, Uma.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 294, No. 6, 01.06.2008.

Research output: Contribution to journalArticle

@article{7617e4e53b4a4f47bbf67f3428e7b1fa,
title = "Functional characterization, localization, and molecular identification of rabbit intestinal N-amino acid transporter",
abstract = "We have characterized the Na-glutamine cotransporter in the rabbit intestinal crypt cell brush border membrane vesicles (BBMV). Substrate specificity experiments showed that crypt cell glutamine uptake is mediated by system N. Real-time PCR experiments showed that SN2 (SLC38A5) mRNA is more abundant in crypt cells compared with SN1 (SLC38A3), indicating that SN2 is the major glutamine transporter present in the apical membrane of the crypt cells. SN2 cDNA was obtained by screening a rabbit intestinal cDNA library with human SN1 used as probe. Rabbit SN2 cDNA encompassed a 473-amino-acid-long open reading frame. SN2 protein displayed 87{\%} identity and 91{\%} similarity to human SN2. Functional characterization studies of rabbit SN2 were performed by using vaccinia virusmediated transient expression system. Substrate specificity of the cloned transporter was identical to that of SN2 described in the literature and matched well with substrate specificity experiments performed using crypt cell BBMV. Cloned rabbit SN2, analogous to its human counterpart, is Li+ tolerant. Hill coefficient for Li+ activation of rabbit SN2-mediated uptake was 1. Taken together, functional data from the crypt cell BBMV and the cloned SN2 cDNA indicate that the crypt cell glutamine transport is most likely mediated by SN2.",
keywords = "Brush border membrane, Na glutamine cotransporter, Small intestine, System N-amino acid transporter",
author = "Talukder, {Jamilur R.} and Ramesh Kekuda and Prosenjit Saha and Prasad, {Puttur D} and Vadivel Ganapathy and Uma Sundaram",
year = "2008",
month = "6",
day = "1",
doi = "10.1152/ajpgi.00578.2007",
language = "English (US)",
volume = "294",
journal = "American journal of physiology. Gastrointestinal and liver physiology",
issn = "0193-1857",
publisher = "American Physiological Society",
number = "6",

}

TY - JOUR

T1 - Functional characterization, localization, and molecular identification of rabbit intestinal N-amino acid transporter

AU - Talukder, Jamilur R.

AU - Kekuda, Ramesh

AU - Saha, Prosenjit

AU - Prasad, Puttur D

AU - Ganapathy, Vadivel

AU - Sundaram, Uma

PY - 2008/6/1

Y1 - 2008/6/1

N2 - We have characterized the Na-glutamine cotransporter in the rabbit intestinal crypt cell brush border membrane vesicles (BBMV). Substrate specificity experiments showed that crypt cell glutamine uptake is mediated by system N. Real-time PCR experiments showed that SN2 (SLC38A5) mRNA is more abundant in crypt cells compared with SN1 (SLC38A3), indicating that SN2 is the major glutamine transporter present in the apical membrane of the crypt cells. SN2 cDNA was obtained by screening a rabbit intestinal cDNA library with human SN1 used as probe. Rabbit SN2 cDNA encompassed a 473-amino-acid-long open reading frame. SN2 protein displayed 87% identity and 91% similarity to human SN2. Functional characterization studies of rabbit SN2 were performed by using vaccinia virusmediated transient expression system. Substrate specificity of the cloned transporter was identical to that of SN2 described in the literature and matched well with substrate specificity experiments performed using crypt cell BBMV. Cloned rabbit SN2, analogous to its human counterpart, is Li+ tolerant. Hill coefficient for Li+ activation of rabbit SN2-mediated uptake was 1. Taken together, functional data from the crypt cell BBMV and the cloned SN2 cDNA indicate that the crypt cell glutamine transport is most likely mediated by SN2.

AB - We have characterized the Na-glutamine cotransporter in the rabbit intestinal crypt cell brush border membrane vesicles (BBMV). Substrate specificity experiments showed that crypt cell glutamine uptake is mediated by system N. Real-time PCR experiments showed that SN2 (SLC38A5) mRNA is more abundant in crypt cells compared with SN1 (SLC38A3), indicating that SN2 is the major glutamine transporter present in the apical membrane of the crypt cells. SN2 cDNA was obtained by screening a rabbit intestinal cDNA library with human SN1 used as probe. Rabbit SN2 cDNA encompassed a 473-amino-acid-long open reading frame. SN2 protein displayed 87% identity and 91% similarity to human SN2. Functional characterization studies of rabbit SN2 were performed by using vaccinia virusmediated transient expression system. Substrate specificity of the cloned transporter was identical to that of SN2 described in the literature and matched well with substrate specificity experiments performed using crypt cell BBMV. Cloned rabbit SN2, analogous to its human counterpart, is Li+ tolerant. Hill coefficient for Li+ activation of rabbit SN2-mediated uptake was 1. Taken together, functional data from the crypt cell BBMV and the cloned SN2 cDNA indicate that the crypt cell glutamine transport is most likely mediated by SN2.

KW - Brush border membrane

KW - Na glutamine cotransporter

KW - Small intestine

KW - System N-amino acid transporter

UR - http://www.scopus.com/inward/record.url?scp=48249131346&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=48249131346&partnerID=8YFLogxK

U2 - 10.1152/ajpgi.00578.2007

DO - 10.1152/ajpgi.00578.2007

M3 - Article

C2 - 18388187

AN - SCOPUS:48249131346

VL - 294

JO - American journal of physiology. Gastrointestinal and liver physiology

JF - American journal of physiology. Gastrointestinal and liver physiology

SN - 0193-1857

IS - 6

ER -