Document Detail


Functional characterization and cloning of amino acid transporter B(0,+) (ATB(0,+)) in primary cultured rat pneumocytes.
MedLine Citation:
PMID:  17960566     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cationic amino acid transport in primary cultured rat pneumocytes exhibiting characteristics of alveolar epithelial type I-like cells are described. Asymmetry and activator ion dependency of (3)H-L-arginine uptake were characterized from the apical or basolateral fluid of pneumocytes grown on permeable support. Substrate specificity of transport was evaluated as a function of (3)H-L-arginine uptake inhibition in the presence of other amino acids. Transepithelial transport studies estimated (3)H-L-arginine flux in the apical-to-basolateral and basolateral-to-apical directions. Full length cDNA of rat amino acid transporter B(0,+) (rATB(0,+)) was cloned and its relative expression level studied. Results indicate that uptake of (3)H-L-arginine from apical fluid is dependent on Na(+) and Cl(-). Zwitterionic and cationic amino acids (excluding L-proline and anionic amino acids) inhibited uptake of (3)H-L-arginine from apical, but not basolateral incubation fluid. Apical-to-basolateral transepithelial flux of (3)H-L-arginine was 20x higher than basolateral-to-apical transport. Kinetic studies of (3)H-L-arginine uptake from apical fluid revealed maximal velocity (V(max)) and Michaelis-Menten constants (K(t)) of 33.32 +/- 2.12 pmol/mg protein/15 min and 0.50 +/- 0.11 mM, respectively, in a cooperative process having a coupling ratio of 1.18 +/- 0.16 with Na(+) and 1.11 +/- 0.13 with Cl(-). Expression of rATB(0,+) mRNA was identified by RT-PCR and Northern analysis. Corresponding cloned 3.2 kb rATB(0,+) cDNA sequence exhibits pronounced homology in deduced amino acid sequence to mouse (95% identity and 97% similarity) and human (89% identity and 95% similarity) ATB(0,+) homologues. We conclude that rat pneumocytes express ATB(0,+), which may partly contribute towards recovering cationic and neutral amino acids from alveolar luminal fluid.
Authors:
Tomomi Uchiyama; Takuya Fujita; Hovhannes J Gukasyan; Kwang-Jin Kim; Zea Borok; Edward D Crandall; Vincent H L Lee
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Journal of cellular physiology     Volume:  214     ISSN:  1097-4652     ISO Abbreviation:  J. Cell. Physiol.     Publication Date:  2008 Mar 
Date Detail:
Created Date:  2007-12-26     Completed Date:  2008-01-18     Revised Date:  2011-11-04    
Medline Journal Info:
Nlm Unique ID:  0050222     Medline TA:  J Cell Physiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  645-54     Citation Subset:  IM    
Copyright Information:
(c) 2007 Wiley-Liss, Inc.
Affiliation:
First Department of Biochemistry, Kyushu University of Health and Welfare, Nobeoka, Miyazaki, Japan.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Amino Acid Transport System ASC / chemistry,  genetics*,  metabolism*
Animals
Arginine / metabolism
Biological Transport
Cells, Cultured
Chlorides
Cloning, Molecular
DNA, Complementary / genetics
Epithelial Cells / metabolism
Gene Expression Regulation
Hydrophobic and Hydrophilic Interactions
Kinetics
Male
Molecular Sequence Data
Pulmonary Alveoli / cytology*,  metabolism*
RNA, Messenger / genetics,  metabolism
Rats
Rats, Sprague-Dawley
Sodium
Substrate Specificity
Time Factors
Tritium / metabolism
Grant Support
ID/Acronym/Agency:
GM59297/GM/NIGMS NIH HHS; HL38578/HL/NHLBI NIH HHS; HL38621/HL/NHLBI NIH HHS; HL38658/HL/NHLBI NIH HHS; HL62569/HL/NHLBI NIH HHS; HL64365/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Amino Acid Transport System ASC; 0/Chlorides; 0/DNA, Complementary; 0/RNA, Messenger; 0/Slc6a14 protein, rat; 10028-17-8/Tritium; 74-79-3/Arginine; 7440-23-5/Sodium

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