Changes in intracellular levels of fructose 2,6-bisphosphate and several glycolytic intermediates in Leishmania major promastigotes as a function of pO2

Leishmania major promastigotes were grown to late log phase, washed and resuspended in Hanks' balanced salt solution, and incubated with glucose at various pO2s in the presence of 5% CO2. Samples were taken at times from 0-40 min and assayed for fructose 2,6-bisphosphate (Fru(2,6)P2), glucose-6...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:Molecular and biochemical parasitology 1991-08, Vol.47 (2), p.161-166
Hauptverfasser: KEEGAN, F. P, BLUM, J. J
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page 166
container_issue 2
container_start_page 161
container_title Molecular and biochemical parasitology
container_volume 47
creator KEEGAN, F. P
BLUM, J. J
description Leishmania major promastigotes were grown to late log phase, washed and resuspended in Hanks' balanced salt solution, and incubated with glucose at various pO2s in the presence of 5% CO2. Samples were taken at times from 0-40 min and assayed for fructose 2,6-bisphosphate (Fru(2,6)P2), glucose-6-phosphate (G6P), fructose-6-phosphate (F6P), phospho(enol)pyruvate (PEP), and ATP. At 95% O2 ATP remained constant throughout the incubation. It did not decrease significantly at 10% O2, but decreased by about 20% and 30% at 6% and 0% O2, respectively. At 95% O2, Fru(2,6)P2 increased about 15-fold within 5 min after the addition of glucose and remained at this high level. At 10%, 6%, and 0% O2 Fru(2,6)P2 rose about 5-fold within 5 min and then declined slightly during the remainder of the incubation. G6P increased from about 0.5 to 12 nmol (mg protein)-1 at 5 min in cells incubated under 95% O2 and then declined to about 5 nmol (mg protein)-1. It increased to about 8 nmol (mg protein)-1 at 5 min and then declined slightly in cells incubated under 10% O2. F6P levels were approximately one-eighth of G6P levels under all conditions, suggesting that phosphohexoseisomerase was not subject to regulation. PEP levels were initially high, but at 95% O2 there was a 50% drop in PEP at 5 min, while at 10%, 6%, and 0% O2 there was less of a decline. The observation that the rise in Fru(2,6)P2 levels at 10%, 6%, or 0% O2 is the same at 5 min and less than the rise at 95% O2 supports the presence of a low affinity oxygen sensor. The different time course of changes in G6P, F6P, and PEP levels suggests that in addition to an activation of pyruvate kinase by Fru(2,6)P2, other regulatory events are also operative at low pO2.
doi_str_mv 10.1016/0166-6851(91)90175-6
format Article
fullrecord <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_72458789</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>72458789</sourcerecordid><originalsourceid>FETCH-LOGICAL-c331t-315b1be6a98ac3c4191d03355523d3468971e3b76d5b86adaac663fe908a9e4e3</originalsourceid><addsrcrecordid>eNpFkd2KFDEQhYO4rOPqGyjkQkTBXpPO_6UM_sHA3uxeh-p0eiZLujMm3cK8iY9r2hlWqCIX9dWpcA5Cbyi5pYTKz7VlI7WgHwz9aAhVopHP0IZq1TaGt_o52jwhL9DLUh4JIUJJeY2uqeGcU75Bf7YHmPa-4DDVmjM4H-MSIePof_tYcBrwkBc3p-Jx-0k2XSjHQ6oNs8cw9bhULkPE-3hyKZ7m4FYhn0ffh8r8U975UA4jTAHwCI8p42NOI5Q57NNKQC08LJObQ5rWi8e79hW6GiAW__ry3qCHb1_vtz-a3d33n9svu8YxRueGUdHRzkswGhxznBraE8aEEC3rGZfaKOpZp2QvOi2hB3BSssEbosF47tkNen_WrV_6tfgy2zGU1QSYfFqKVS0XWmlTQX4GXU6lZD_YYw4j5JOlxK6B2NVtu7ptTa01ECvr2tuL_tJVS_4vnROo83eXORQHccgwuVCeMEGUUKplfwEMg5W_</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>72458789</pqid></control><display><type>article</type><title>Changes in intracellular levels of fructose 2,6-bisphosphate and several glycolytic intermediates in Leishmania major promastigotes as a function of pO2</title><source>MEDLINE</source><source>Access via ScienceDirect (Elsevier)</source><creator>KEEGAN, F. P ; BLUM, J. J</creator><creatorcontrib>KEEGAN, F. P ; BLUM, J. J</creatorcontrib><description>Leishmania major promastigotes were grown to late log phase, washed and resuspended in Hanks' balanced salt solution, and incubated with glucose at various pO2s in the presence of 5% CO2. Samples were taken at times from 0-40 min and assayed for fructose 2,6-bisphosphate (Fru(2,6)P2), glucose-6-phosphate (G6P), fructose-6-phosphate (F6P), phospho(enol)pyruvate (PEP), and ATP. At 95% O2 ATP remained constant throughout the incubation. It did not decrease significantly at 10% O2, but decreased by about 20% and 30% at 6% and 0% O2, respectively. At 95% O2, Fru(2,6)P2 increased about 15-fold within 5 min after the addition of glucose and remained at this high level. At 10%, 6%, and 0% O2 Fru(2,6)P2 rose about 5-fold within 5 min and then declined slightly during the remainder of the incubation. G6P increased from about 0.5 to 12 nmol (mg protein)-1 at 5 min in cells incubated under 95% O2 and then declined to about 5 nmol (mg protein)-1. It increased to about 8 nmol (mg protein)-1 at 5 min and then declined slightly in cells incubated under 10% O2. F6P levels were approximately one-eighth of G6P levels under all conditions, suggesting that phosphohexoseisomerase was not subject to regulation. PEP levels were initially high, but at 95% O2 there was a 50% drop in PEP at 5 min, while at 10%, 6%, and 0% O2 there was less of a decline. The observation that the rise in Fru(2,6)P2 levels at 10%, 6%, or 0% O2 is the same at 5 min and less than the rise at 95% O2 supports the presence of a low affinity oxygen sensor. The different time course of changes in G6P, F6P, and PEP levels suggests that in addition to an activation of pyruvate kinase by Fru(2,6)P2, other regulatory events are also operative at low pO2.</description><identifier>ISSN: 0166-6851</identifier><identifier>EISSN: 1872-9428</identifier><identifier>DOI: 10.1016/0166-6851(91)90175-6</identifier><identifier>PMID: 1944414</identifier><identifier>CODEN: MBIPDP</identifier><language>eng</language><publisher>Shannon: Elsevier Science</publisher><subject>Adenosine Triphosphate - metabolism ; Anaerobiosis ; Animals ; Biochemistry. Physiology. Immunology. Molecular biology ; Biological and medical sciences ; Fructosediphosphates - metabolism ; Fructosephosphates - metabolism ; Fundamental and applied biological sciences. Psychology ; Glucose - metabolism ; Glucose-6-Phosphate ; Glucosephosphates - metabolism ; Glycolysis ; Kinetics ; Leishmania tropica - growth &amp; development ; Leishmania tropica - metabolism ; Oxygen - metabolism ; Phosphoenolpyruvate - metabolism ; Protozoa ; Tropical medicine</subject><ispartof>Molecular and biochemical parasitology, 1991-08, Vol.47 (2), p.161-166</ispartof><rights>1992 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c331t-315b1be6a98ac3c4191d03355523d3468971e3b76d5b86adaac663fe908a9e4e3</citedby><cites>FETCH-LOGICAL-c331t-315b1be6a98ac3c4191d03355523d3468971e3b76d5b86adaac663fe908a9e4e3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&amp;idt=5075772$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1944414$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>KEEGAN, F. P</creatorcontrib><creatorcontrib>BLUM, J. J</creatorcontrib><title>Changes in intracellular levels of fructose 2,6-bisphosphate and several glycolytic intermediates in Leishmania major promastigotes as a function of pO2</title><title>Molecular and biochemical parasitology</title><addtitle>Mol Biochem Parasitol</addtitle><description>Leishmania major promastigotes were grown to late log phase, washed and resuspended in Hanks' balanced salt solution, and incubated with glucose at various pO2s in the presence of 5% CO2. Samples were taken at times from 0-40 min and assayed for fructose 2,6-bisphosphate (Fru(2,6)P2), glucose-6-phosphate (G6P), fructose-6-phosphate (F6P), phospho(enol)pyruvate (PEP), and ATP. At 95% O2 ATP remained constant throughout the incubation. It did not decrease significantly at 10% O2, but decreased by about 20% and 30% at 6% and 0% O2, respectively. At 95% O2, Fru(2,6)P2 increased about 15-fold within 5 min after the addition of glucose and remained at this high level. At 10%, 6%, and 0% O2 Fru(2,6)P2 rose about 5-fold within 5 min and then declined slightly during the remainder of the incubation. G6P increased from about 0.5 to 12 nmol (mg protein)-1 at 5 min in cells incubated under 95% O2 and then declined to about 5 nmol (mg protein)-1. It increased to about 8 nmol (mg protein)-1 at 5 min and then declined slightly in cells incubated under 10% O2. F6P levels were approximately one-eighth of G6P levels under all conditions, suggesting that phosphohexoseisomerase was not subject to regulation. PEP levels were initially high, but at 95% O2 there was a 50% drop in PEP at 5 min, while at 10%, 6%, and 0% O2 there was less of a decline. The observation that the rise in Fru(2,6)P2 levels at 10%, 6%, or 0% O2 is the same at 5 min and less than the rise at 95% O2 supports the presence of a low affinity oxygen sensor. The different time course of changes in G6P, F6P, and PEP levels suggests that in addition to an activation of pyruvate kinase by Fru(2,6)P2, other regulatory events are also operative at low pO2.</description><subject>Adenosine Triphosphate - metabolism</subject><subject>Anaerobiosis</subject><subject>Animals</subject><subject>Biochemistry. Physiology. Immunology. Molecular biology</subject><subject>Biological and medical sciences</subject><subject>Fructosediphosphates - metabolism</subject><subject>Fructosephosphates - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Glucose - metabolism</subject><subject>Glucose-6-Phosphate</subject><subject>Glucosephosphates - metabolism</subject><subject>Glycolysis</subject><subject>Kinetics</subject><subject>Leishmania tropica - growth &amp; development</subject><subject>Leishmania tropica - metabolism</subject><subject>Oxygen - metabolism</subject><subject>Phosphoenolpyruvate - metabolism</subject><subject>Protozoa</subject><subject>Tropical medicine</subject><issn>0166-6851</issn><issn>1872-9428</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1991</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkd2KFDEQhYO4rOPqGyjkQkTBXpPO_6UM_sHA3uxeh-p0eiZLujMm3cK8iY9r2hlWqCIX9dWpcA5Cbyi5pYTKz7VlI7WgHwz9aAhVopHP0IZq1TaGt_o52jwhL9DLUh4JIUJJeY2uqeGcU75Bf7YHmPa-4DDVmjM4H-MSIePof_tYcBrwkBc3p-Jx-0k2XSjHQ6oNs8cw9bhULkPE-3hyKZ7m4FYhn0ffh8r8U975UA4jTAHwCI8p42NOI5Q57NNKQC08LJObQ5rWi8e79hW6GiAW__ry3qCHb1_vtz-a3d33n9svu8YxRueGUdHRzkswGhxznBraE8aEEC3rGZfaKOpZp2QvOi2hB3BSssEbosF47tkNen_WrV_6tfgy2zGU1QSYfFqKVS0XWmlTQX4GXU6lZD_YYw4j5JOlxK6B2NVtu7ptTa01ECvr2tuL_tJVS_4vnROo83eXORQHccgwuVCeMEGUUKplfwEMg5W_</recordid><startdate>19910801</startdate><enddate>19910801</enddate><creator>KEEGAN, F. P</creator><creator>BLUM, J. J</creator><general>Elsevier 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>7X8</scope></search><sort><creationdate>19910801</creationdate><title>Changes in intracellular levels of fructose 2,6-bisphosphate and several glycolytic intermediates in Leishmania major promastigotes as a function of pO2</title><author>KEEGAN, F. P ; BLUM, J. J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c331t-315b1be6a98ac3c4191d03355523d3468971e3b76d5b86adaac663fe908a9e4e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1991</creationdate><topic>Adenosine Triphosphate - metabolism</topic><topic>Anaerobiosis</topic><topic>Animals</topic><topic>Biochemistry. Physiology. Immunology. Molecular biology</topic><topic>Biological and medical sciences</topic><topic>Fructosediphosphates - metabolism</topic><topic>Fructosephosphates - metabolism</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Glucose - metabolism</topic><topic>Glucose-6-Phosphate</topic><topic>Glucosephosphates - metabolism</topic><topic>Glycolysis</topic><topic>Kinetics</topic><topic>Leishmania tropica - growth &amp; development</topic><topic>Leishmania tropica - metabolism</topic><topic>Oxygen - metabolism</topic><topic>Phosphoenolpyruvate - metabolism</topic><topic>Protozoa</topic><topic>Tropical medicine</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>KEEGAN, F. P</creatorcontrib><creatorcontrib>BLUM, J. J</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>MEDLINE - Academic</collection><jtitle>Molecular and biochemical parasitology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>KEEGAN, F. P</au><au>BLUM, J. J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Changes in intracellular levels of fructose 2,6-bisphosphate and several glycolytic intermediates in Leishmania major promastigotes as a function of pO2</atitle><jtitle>Molecular and biochemical parasitology</jtitle><addtitle>Mol Biochem Parasitol</addtitle><date>1991-08-01</date><risdate>1991</risdate><volume>47</volume><issue>2</issue><spage>161</spage><epage>166</epage><pages>161-166</pages><issn>0166-6851</issn><eissn>1872-9428</eissn><coden>MBIPDP</coden><abstract>Leishmania major promastigotes were grown to late log phase, washed and resuspended in Hanks' balanced salt solution, and incubated with glucose at various pO2s in the presence of 5% CO2. Samples were taken at times from 0-40 min and assayed for fructose 2,6-bisphosphate (Fru(2,6)P2), glucose-6-phosphate (G6P), fructose-6-phosphate (F6P), phospho(enol)pyruvate (PEP), and ATP. At 95% O2 ATP remained constant throughout the incubation. It did not decrease significantly at 10% O2, but decreased by about 20% and 30% at 6% and 0% O2, respectively. At 95% O2, Fru(2,6)P2 increased about 15-fold within 5 min after the addition of glucose and remained at this high level. At 10%, 6%, and 0% O2 Fru(2,6)P2 rose about 5-fold within 5 min and then declined slightly during the remainder of the incubation. G6P increased from about 0.5 to 12 nmol (mg protein)-1 at 5 min in cells incubated under 95% O2 and then declined to about 5 nmol (mg protein)-1. It increased to about 8 nmol (mg protein)-1 at 5 min and then declined slightly in cells incubated under 10% O2. F6P levels were approximately one-eighth of G6P levels under all conditions, suggesting that phosphohexoseisomerase was not subject to regulation. PEP levels were initially high, but at 95% O2 there was a 50% drop in PEP at 5 min, while at 10%, 6%, and 0% O2 there was less of a decline. The observation that the rise in Fru(2,6)P2 levels at 10%, 6%, or 0% O2 is the same at 5 min and less than the rise at 95% O2 supports the presence of a low affinity oxygen sensor. The different time course of changes in G6P, F6P, and PEP levels suggests that in addition to an activation of pyruvate kinase by Fru(2,6)P2, other regulatory events are also operative at low pO2.</abstract><cop>Shannon</cop><pub>Elsevier Science</pub><pmid>1944414</pmid><doi>10.1016/0166-6851(91)90175-6</doi><tpages>6</tpages></addata></record>
fulltext fulltext
identifier ISSN: 0166-6851
ispartof Molecular and biochemical parasitology, 1991-08, Vol.47 (2), p.161-166
issn 0166-6851
1872-9428
language eng
recordid cdi_proquest_miscellaneous_72458789
source MEDLINE; Access via ScienceDirect (Elsevier)
subjects Adenosine Triphosphate - metabolism
Anaerobiosis
Animals
Biochemistry. Physiology. Immunology. Molecular biology
Biological and medical sciences
Fructosediphosphates - metabolism
Fructosephosphates - metabolism
Fundamental and applied biological sciences. Psychology
Glucose - metabolism
Glucose-6-Phosphate
Glucosephosphates - metabolism
Glycolysis
Kinetics
Leishmania tropica - growth & development
Leishmania tropica - metabolism
Oxygen - metabolism
Phosphoenolpyruvate - metabolism
Protozoa
Tropical medicine
title Changes in intracellular levels of fructose 2,6-bisphosphate and several glycolytic intermediates in Leishmania major promastigotes as a function of pO2
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T15%3A14%3A35IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Changes%20in%20intracellular%20levels%20of%20fructose%202,6-bisphosphate%20and%20several%20glycolytic%20intermediates%20in%20Leishmania%20major%20promastigotes%20as%20a%20function%20of%20pO2&rft.jtitle=Molecular%20and%20biochemical%20parasitology&rft.au=KEEGAN,%20F.%20P&rft.date=1991-08-01&rft.volume=47&rft.issue=2&rft.spage=161&rft.epage=166&rft.pages=161-166&rft.issn=0166-6851&rft.eissn=1872-9428&rft.coden=MBIPDP&rft_id=info:doi/10.1016/0166-6851(91)90175-6&rft_dat=%3Cproquest_cross%3E72458789%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=72458789&rft_id=info:pmid/1944414&rfr_iscdi=true