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Ammonia permeability of the aquaglyceroporins from Plasmodium falciparum, Toxoplasma gondii and Trypansoma brucei
Summary Plasmodium falciparum uses amino acids from haemoglobin degradation mainly for protein biosynthesis. Glutamine, however, is mostly oxidized to 2‐oxoglutarate to restore NAD(P)H + H+. In this process two molecules of ammonia are released. We determined an ammonia production of 9 mmol h−1 per...
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Published in: | Molecular microbiology 2006-09, Vol.61 (6), p.1598-1608 |
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creator | Zeuthen, Thomas Wu, Binghua Pavlovic‐Djuranovic, Slavica Holm, Lars M. Uzcategui, Nestor L. Duszenko, Michael Kun, Jürgen F. J. Schultz, Joachim E. Beitz, Eric |
description | Summary
Plasmodium falciparum uses amino acids from haemoglobin degradation mainly for protein biosynthesis. Glutamine, however, is mostly oxidized to 2‐oxoglutarate to restore NAD(P)H + H+. In this process two molecules of ammonia are released. We determined an ammonia production of 9 mmol h−1 per litre of infected red blood cells in the early trophozoite stage. External application of ammonia yielded a cytotoxic IC50 concentration of 2.8 mM. As plasmodia cannot metabolize ammonia it must be exported. Yet, no biochemical or genomic evidences exist that plasmodia possess classical ammonium transporters. We expressed the P. falciparum aquaglyceroporin (PfAQP) in Xenopus laevis oocytes and examined whether it may serve as an exit pathway for ammonia. We show that injected oocytes: (i) acidify the medium due to ammonia uptake, (ii) take up [14C]methylamine and [14C]formamide, (iii) swell in solution with formamide and acetamide and (iv) display an ammonia‐induced NH4+‐dependent clamp current. Further, a yeast strain lacking the endogenous aquaglyceroporin (Fps1) is rescued by expression of PfAQP which provides for the efflux of toxic methylamine. Ammonia permeability was similarly established for the aquaglyceroporins from Toxoplasma gondii and Trypanosoma brucei. Apparently, these aquaglyceroporins are important for the release of ammonia derived from amino acid breakdown. |
doi_str_mv | 10.1111/j.1365-2958.2006.05325.x |
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Plasmodium falciparum uses amino acids from haemoglobin degradation mainly for protein biosynthesis. Glutamine, however, is mostly oxidized to 2‐oxoglutarate to restore NAD(P)H + H+. In this process two molecules of ammonia are released. We determined an ammonia production of 9 mmol h−1 per litre of infected red blood cells in the early trophozoite stage. External application of ammonia yielded a cytotoxic IC50 concentration of 2.8 mM. As plasmodia cannot metabolize ammonia it must be exported. Yet, no biochemical or genomic evidences exist that plasmodia possess classical ammonium transporters. We expressed the P. falciparum aquaglyceroporin (PfAQP) in Xenopus laevis oocytes and examined whether it may serve as an exit pathway for ammonia. We show that injected oocytes: (i) acidify the medium due to ammonia uptake, (ii) take up [14C]methylamine and [14C]formamide, (iii) swell in solution with formamide and acetamide and (iv) display an ammonia‐induced NH4+‐dependent clamp current. Further, a yeast strain lacking the endogenous aquaglyceroporin (Fps1) is rescued by expression of PfAQP which provides for the efflux of toxic methylamine. Ammonia permeability was similarly established for the aquaglyceroporins from Toxoplasma gondii and Trypanosoma brucei. Apparently, these aquaglyceroporins are important for the release of ammonia derived from amino acid breakdown.</description><identifier>ISSN: 0950-382X</identifier><identifier>EISSN: 1365-2958</identifier><identifier>DOI: 10.1111/j.1365-2958.2006.05325.x</identifier><identifier>PMID: 16889642</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Amino acids ; Ammonia ; Ammonia - metabolism ; Animals ; Aquaglyceroporins - genetics ; Aquaglyceroporins - physiology ; Biological and medical sciences ; Fundamental and applied biological sciences. Psychology ; Genetic Complementation Test ; Hemoglobin ; Membrane Proteins - genetics ; Membrane Proteins - physiology ; Methylamines - toxicity ; Microbiology ; Oocytes ; Parasitic protozoa ; Permeability ; Plasmodium falciparum ; Plasmodium falciparum - growth & development ; Plasmodium falciparum - metabolism ; Proteins ; Saccharomyces cerevisiae - genetics ; Saccharomyces cerevisiae - physiology ; Saccharomyces cerevisiae Proteins - genetics ; Saccharomyces cerevisiae Proteins - physiology ; Toxoplasma - growth & development ; Toxoplasma - metabolism ; Toxoplasma gondii ; Trypanosoma brucei ; Trypanosoma brucei brucei - growth & development ; Trypanosoma brucei brucei - metabolism ; Xenopus laevis</subject><ispartof>Molecular microbiology, 2006-09, Vol.61 (6), p.1598-1608</ispartof><rights>2006 INIST-CNRS</rights><rights>Copyright Blackwell Publishing Sep 2006</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5715-2078e821fd1ce95c1c5c40cd94de177927c5d8009bc0ea1f7ba76ffe0a1dc93d3</citedby><cites>FETCH-LOGICAL-c5715-2078e821fd1ce95c1c5c40cd94de177927c5d8009bc0ea1f7ba76ffe0a1dc93d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27923,27924</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=18090051$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16889642$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zeuthen, Thomas</creatorcontrib><creatorcontrib>Wu, Binghua</creatorcontrib><creatorcontrib>Pavlovic‐Djuranovic, Slavica</creatorcontrib><creatorcontrib>Holm, Lars M.</creatorcontrib><creatorcontrib>Uzcategui, Nestor L.</creatorcontrib><creatorcontrib>Duszenko, Michael</creatorcontrib><creatorcontrib>Kun, Jürgen F. J.</creatorcontrib><creatorcontrib>Schultz, Joachim E.</creatorcontrib><creatorcontrib>Beitz, Eric</creatorcontrib><title>Ammonia permeability of the aquaglyceroporins from Plasmodium falciparum, Toxoplasma gondii and Trypansoma brucei</title><title>Molecular microbiology</title><addtitle>Mol Microbiol</addtitle><description>Summary
Plasmodium falciparum uses amino acids from haemoglobin degradation mainly for protein biosynthesis. Glutamine, however, is mostly oxidized to 2‐oxoglutarate to restore NAD(P)H + H+. In this process two molecules of ammonia are released. We determined an ammonia production of 9 mmol h−1 per litre of infected red blood cells in the early trophozoite stage. External application of ammonia yielded a cytotoxic IC50 concentration of 2.8 mM. As plasmodia cannot metabolize ammonia it must be exported. Yet, no biochemical or genomic evidences exist that plasmodia possess classical ammonium transporters. We expressed the P. falciparum aquaglyceroporin (PfAQP) in Xenopus laevis oocytes and examined whether it may serve as an exit pathway for ammonia. We show that injected oocytes: (i) acidify the medium due to ammonia uptake, (ii) take up [14C]methylamine and [14C]formamide, (iii) swell in solution with formamide and acetamide and (iv) display an ammonia‐induced NH4+‐dependent clamp current. Further, a yeast strain lacking the endogenous aquaglyceroporin (Fps1) is rescued by expression of PfAQP which provides for the efflux of toxic methylamine. Ammonia permeability was similarly established for the aquaglyceroporins from Toxoplasma gondii and Trypanosoma brucei. Apparently, these aquaglyceroporins are important for the release of ammonia derived from amino acid breakdown.</description><subject>Amino acids</subject><subject>Ammonia</subject><subject>Ammonia - metabolism</subject><subject>Animals</subject><subject>Aquaglyceroporins - genetics</subject><subject>Aquaglyceroporins - physiology</subject><subject>Biological and medical sciences</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genetic Complementation Test</subject><subject>Hemoglobin</subject><subject>Membrane Proteins - genetics</subject><subject>Membrane Proteins - physiology</subject><subject>Methylamines - toxicity</subject><subject>Microbiology</subject><subject>Oocytes</subject><subject>Parasitic protozoa</subject><subject>Permeability</subject><subject>Plasmodium falciparum</subject><subject>Plasmodium falciparum - growth & development</subject><subject>Plasmodium falciparum - metabolism</subject><subject>Proteins</subject><subject>Saccharomyces cerevisiae - genetics</subject><subject>Saccharomyces cerevisiae - physiology</subject><subject>Saccharomyces cerevisiae Proteins - genetics</subject><subject>Saccharomyces cerevisiae Proteins - physiology</subject><subject>Toxoplasma - growth & development</subject><subject>Toxoplasma - metabolism</subject><subject>Toxoplasma gondii</subject><subject>Trypanosoma brucei</subject><subject>Trypanosoma brucei brucei - growth & development</subject><subject>Trypanosoma brucei brucei - metabolism</subject><subject>Xenopus laevis</subject><issn>0950-382X</issn><issn>1365-2958</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><recordid>eNqNkU-PFCEQxYnRuOPqVzDERE9OW8DQDQcPm41_NtmNHsbEG6GBXpk0TQ9MZ6e_vbQzcRNPcoFU_V7lUQ8hTKAi5XzYVYTVfE0lFxUFqCvgjPLq-ASt_jaeohVIDmsm6M8L9CLnHQBhULPn6ILUQsh6Q1dofxVCHLzGo0vB6db3_jDj2OHDL4f1ftL3_WxcimNMfsi4SzHg773OIVo_Bdzp3vhRpym8x9t4jOPS0vg-DtZ7rAeLt2ke9ZBjqbZpMs6_RM-KKrtX5_sS_fj8aXv9dX377cvN9dXt2vCGlC9AI5ygpLPEOMkNMdxswFi5sY40jaSN4VYAyNaA06RrWt3UXedAE2sks-wSvTvNHVPcTy4fVPDZuL7Xg4tTVkRy1gCnBXzzD7iLUxqKt8LUnIGkpEDiBJkUc06uU2PyQadZEVBLJmqnltWrZfVqyUT9yUQdi_T1ef7UBmcfhecQCvD2DOhsdN8lPRifHzkBEoAvHj6euAffu_m_Dai7u5vlxX4DQr6pwg</recordid><startdate>200609</startdate><enddate>200609</enddate><creator>Zeuthen, Thomas</creator><creator>Wu, Binghua</creator><creator>Pavlovic‐Djuranovic, Slavica</creator><creator>Holm, Lars M.</creator><creator>Uzcategui, Nestor L.</creator><creator>Duszenko, Michael</creator><creator>Kun, Jürgen F. J.</creator><creator>Schultz, Joachim E.</creator><creator>Beitz, Eric</creator><general>Blackwell Publishing Ltd</general><general>Blackwell Science</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>200609</creationdate><title>Ammonia permeability of the aquaglyceroporins from Plasmodium falciparum, Toxoplasma gondii and Trypansoma brucei</title><author>Zeuthen, Thomas ; Wu, Binghua ; Pavlovic‐Djuranovic, Slavica ; Holm, Lars M. ; Uzcategui, Nestor L. ; Duszenko, Michael ; Kun, Jürgen F. 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Psychology</topic><topic>Genetic Complementation Test</topic><topic>Hemoglobin</topic><topic>Membrane Proteins - genetics</topic><topic>Membrane Proteins - physiology</topic><topic>Methylamines - toxicity</topic><topic>Microbiology</topic><topic>Oocytes</topic><topic>Parasitic protozoa</topic><topic>Permeability</topic><topic>Plasmodium falciparum</topic><topic>Plasmodium falciparum - growth & development</topic><topic>Plasmodium falciparum - metabolism</topic><topic>Proteins</topic><topic>Saccharomyces cerevisiae - genetics</topic><topic>Saccharomyces cerevisiae - physiology</topic><topic>Saccharomyces cerevisiae Proteins - genetics</topic><topic>Saccharomyces cerevisiae Proteins - physiology</topic><topic>Toxoplasma - growth & development</topic><topic>Toxoplasma - metabolism</topic><topic>Toxoplasma gondii</topic><topic>Trypanosoma brucei</topic><topic>Trypanosoma brucei brucei - growth & development</topic><topic>Trypanosoma brucei brucei - metabolism</topic><topic>Xenopus laevis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zeuthen, Thomas</creatorcontrib><creatorcontrib>Wu, Binghua</creatorcontrib><creatorcontrib>Pavlovic‐Djuranovic, Slavica</creatorcontrib><creatorcontrib>Holm, Lars M.</creatorcontrib><creatorcontrib>Uzcategui, Nestor L.</creatorcontrib><creatorcontrib>Duszenko, Michael</creatorcontrib><creatorcontrib>Kun, Jürgen F. J.</creatorcontrib><creatorcontrib>Schultz, Joachim E.</creatorcontrib><creatorcontrib>Beitz, Eric</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Molecular microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zeuthen, Thomas</au><au>Wu, Binghua</au><au>Pavlovic‐Djuranovic, Slavica</au><au>Holm, Lars M.</au><au>Uzcategui, Nestor L.</au><au>Duszenko, Michael</au><au>Kun, Jürgen F. J.</au><au>Schultz, Joachim E.</au><au>Beitz, Eric</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Ammonia permeability of the aquaglyceroporins from Plasmodium falciparum, Toxoplasma gondii and Trypansoma brucei</atitle><jtitle>Molecular microbiology</jtitle><addtitle>Mol Microbiol</addtitle><date>2006-09</date><risdate>2006</risdate><volume>61</volume><issue>6</issue><spage>1598</spage><epage>1608</epage><pages>1598-1608</pages><issn>0950-382X</issn><eissn>1365-2958</eissn><abstract>Summary
Plasmodium falciparum uses amino acids from haemoglobin degradation mainly for protein biosynthesis. Glutamine, however, is mostly oxidized to 2‐oxoglutarate to restore NAD(P)H + H+. In this process two molecules of ammonia are released. We determined an ammonia production of 9 mmol h−1 per litre of infected red blood cells in the early trophozoite stage. External application of ammonia yielded a cytotoxic IC50 concentration of 2.8 mM. As plasmodia cannot metabolize ammonia it must be exported. Yet, no biochemical or genomic evidences exist that plasmodia possess classical ammonium transporters. We expressed the P. falciparum aquaglyceroporin (PfAQP) in Xenopus laevis oocytes and examined whether it may serve as an exit pathway for ammonia. We show that injected oocytes: (i) acidify the medium due to ammonia uptake, (ii) take up [14C]methylamine and [14C]formamide, (iii) swell in solution with formamide and acetamide and (iv) display an ammonia‐induced NH4+‐dependent clamp current. Further, a yeast strain lacking the endogenous aquaglyceroporin (Fps1) is rescued by expression of PfAQP which provides for the efflux of toxic methylamine. Ammonia permeability was similarly established for the aquaglyceroporins from Toxoplasma gondii and Trypanosoma brucei. Apparently, these aquaglyceroporins are important for the release of ammonia derived from amino acid breakdown.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>16889642</pmid><doi>10.1111/j.1365-2958.2006.05325.x</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Amino acids Ammonia Ammonia - metabolism Animals Aquaglyceroporins - genetics Aquaglyceroporins - physiology Biological and medical sciences Fundamental and applied biological sciences. Psychology Genetic Complementation Test Hemoglobin Membrane Proteins - genetics Membrane Proteins - physiology Methylamines - toxicity Microbiology Oocytes Parasitic protozoa Permeability Plasmodium falciparum Plasmodium falciparum - growth & development Plasmodium falciparum - metabolism Proteins Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - physiology Saccharomyces cerevisiae Proteins - genetics Saccharomyces cerevisiae Proteins - physiology Toxoplasma - growth & development Toxoplasma - metabolism Toxoplasma gondii Trypanosoma brucei Trypanosoma brucei brucei - growth & development Trypanosoma brucei brucei - metabolism Xenopus laevis |
title | Ammonia permeability of the aquaglyceroporins from Plasmodium falciparum, Toxoplasma gondii and Trypansoma brucei |
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