| Effect of human carbonic anhydrase II on the activity of the human electrogenic Na/HCO3 cotransporter NBCe1-A in Xenopus oocytes. | |
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MedLine Citation:
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PMID: 16687407 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Others report that carbonic anhydrase II (CA II) binds to the C termini of the anion exchanger AE1 and the electrogenic Na/HCO3 cotransporter NBCe1-A, enhancing transport. After injecting oocytes with NBCe1-A cRNA (Day 0), we measured NBC current (I(NBC)) by two-electrode voltage clamp (Day 3), injected CA II protein + Tris or just Tris (Day 3), measured I(NBC) or the initial rate at which the intracellular pH fell (dpH(i)/dt) upon applying 5% CO2 (Day 4), exposed oocytes to the permeant CA inhibitor ethoxzolamide (EZA), and measured I(NBC) or dpH(i)/dt (Day 4). Because dpH(i)/dt was greater in CA II than Tris oocytes, and EZA eliminated the difference, injected CA II was functional. I(NBC) slope conductance was unaffected by injecting CA II. Moreover, EZA had identical effects in CA II versus Tris oocytes. Thus, injected CA II does not enhance NBC activity. In a second protocol, we made a fusion protein with enhanced green fluorescent protein (EGFP) at the 5' end of NBCe1-A and CA II at the 3' end (EGFP-e1-CAII). We measured I(NBC) or dpH(i)/dt (days 3-4), exposed oocytes to EZA, and measured I(NBC) or dpH(i)/dt (Day 3-4). dpH(i)/dt was greater in oocytes expressing EGFP-e1-CA II versus EGFP-e1, and EZA eliminated the difference. Thus, fused CA II was functional. Slope conductances of EGFP-e1-CAII versus EGFP-e1 oocytes were indistinguishable, and EZA had no effect. Thus, even when fused to NBCe1-A, CA II does not enhance NBCe1-A activity. |
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Authors:
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Jing Lu; Christopher M Daly; Mark D Parker; Harindarpal S Gill; Peter M Piermarini; Marc F Pelletier; Walter F Boron |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't Date: 2006-05-10 |
Journal Detail:
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Title: The Journal of biological chemistry Volume: 281 ISSN: 0021-9258 ISO Abbreviation: J. Biol. Chem. Publication Date: 2006 Jul |
Date Detail:
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Created Date: 2006-07-10 Completed Date: 2006-08-21 Revised Date: 2012-11-16 |
Medline Journal Info:
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Nlm Unique ID: 2985121R Medline TA: J Biol Chem Country: United States |
Other Details:
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Languages: eng Pagination: 19241-50 Citation Subset: IM |
Affiliation:
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Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut 06520, USA. Jing.Lu@Yale.edu |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Carbonic Anhydrase II / chemistry* Electrophysiology Ethoxzolamide / pharmacology Green Fluorescent Proteins / metabolism Humans Hydrogen-Ion Concentration Models, Biological Oocytes / metabolism* RNA, Complementary / metabolism Sodium-Bicarbonate Symporters / metabolism*, physiology Time Factors Xenopus / metabolism* |
| Grant Support | |
ID/Acronym/Agency:
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DK-30344/DK/NIDDK NIH HHS; F32 DK075258/DK/NIDDK NIH HHS; F32 DK075258-03/DK/NIDDK NIH HHS; K01 DK082646/DK/NIDDK NIH HHS; P30-34989//PHS HHS |
| Chemical | |
Reg. No./Substance:
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0/RNA, Complementary; 0/SLC4A4 protein, human; 0/Sodium-Bicarbonate Symporters; 147336-22-9/Green Fluorescent Proteins; 452-35-7/Ethoxzolamide; EC 4.2.1.-/Carbonic Anhydrase II |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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