| Functional characterization and cloning of amino acid transporter B(0,+) (ATB(0,+)) in primary cultured rat pneumocytes. | |
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MedLine Citation:
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PMID: 17960566 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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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:
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Type: Journal Article; Research Support, N.I.H., Extramural |
Journal Detail:
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Title: Journal of cellular physiology Volume: 214 ISSN: 1097-4652 ISO Abbreviation: J. Cell. Physiol. Publication Date: 2008 Mar |
Date Detail:
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Created Date: 2007-12-26 Completed Date: 2008-01-18 Revised Date: 2011-11-04 |
Medline Journal Info:
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Nlm Unique ID: 0050222 Medline TA: J Cell Physiol Country: United States |
Other Details:
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Languages: eng Pagination: 645-54 Citation Subset: IM |
Copyright Information:
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(c) 2007 Wiley-Liss, Inc. |
Affiliation:
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First Department of Biochemistry, Kyushu University of Health and Welfare, Nobeoka, Miyazaki, Japan. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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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:
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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:
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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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