A role for gamma-glutamyl transpeptidase and the amino acid transport system xc- in cystine transport by a human pancreatic duct cell line
1. The roles of the gamma-glutamyl cycle and the anionic amino acid transport system xc- in mediating L-cystine uptake were investigated in cultured human pancreatic duct PaTu 8902 cells. This cell line exhibits morphological features of normal pancreatic duct cells and expresses gamma-glutamyl tran...
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| Veröffentlicht in: | The Journal of physiology 1995-05, Vol.485 (Pt 1), p.167-177 |
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| creator | J H Sweiry J Sastre J Viña H P Elsässer G E Mann |
| description | 1. The roles of the gamma-glutamyl cycle and the anionic amino acid transport system xc- in mediating L-cystine uptake were
investigated in cultured human pancreatic duct PaTu 8902 cells. This cell line exhibits morphological features of normal pancreatic
duct cells and expresses gamma-glutamyl transpeptidase (gamma-GT, EC 2.3.2.2), an enzyme involved in the metabolism and regulation
of intracellular glutathione (GSH). 2. Uptake of L-cystine (10 microM) was linear for up to 10 min, temperature dependent,
Na+ independent, saturable (Michaelis-Menten constant (Km), 86 +/- 25 microM; maximal velocity (Vmax), 109 +/- 33 nmol (mg
protein)-1 h-1) and reduced by 80-90% by a 50-fold excess concentration of L-glutamate and L-homocysteic acid, but not L-aspartate.
These transport properties resemble those described for system xc-, which exchanges cystine for intracellular glutamate. 3.
Acivicin, a known inhibitor of gamma-GT, decreased gamma-GT activity from 2.58 +/- 0.96 to 0.97 +/- 0.11 mU (mg protein)-1
and decreased the initial rates of L-cystine and L-glutamine uptake by 41-55%. Anthglutin (1-gamma-L-glutamyl-2-(2-carboxyphenylhyl)hydrazine),
a structurally different inhibitor of gamma-GT, also caused a concentration-dependent (0.01-1 mM) decrease in gamma-GT activity
and L-cystine uptake. 4. Neither acivicin nor anthglutin inhibited the uptake of L-glutamate, a poor substrate for gamma-GT.
5. In the presence of a 500-fold excess concentration of glutamate, which should abolish entry of cystine via system xc-,
the remaining fraction of cystine transport was inhibited by 50% by acivicin, suggesting that transport is, in part, dependent
on the activity of gamma-GT. 6. Cystine transport was also 60-80% inhibited by a series of gamma-glutamyl amino acids (5 mM)
including gamma-glutamyl-glutamate, gamma-glutamyl-glutamine and gamma-glutamyl-glycine. alpha-Dipeptides inhibited cystine
transport by only 6-22%. 7. These findings demonstrate that in human pancreatic duct PaTu 8902 cells, cystine uptake is mediated
by system xc- (50-60%) and the gamma-glutamyl cycle. Our results provide the first evidence linking gamma-GT with cystine
transport in human epithelial cells and are of relevance in view of the importance of cystine as a sulphur amino acid source
for GSH synthesis in cells exposed to oxidative stress. |
| doi_str_mv | 10.1113/jphysiol.1995.sp020721 |
| format | Article |
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investigated in cultured human pancreatic duct PaTu 8902 cells. This cell line exhibits morphological features of normal pancreatic
duct cells and expresses gamma-glutamyl transpeptidase (gamma-GT, EC 2.3.2.2), an enzyme involved in the metabolism and regulation
of intracellular glutathione (GSH). 2. Uptake of L-cystine (10 microM) was linear for up to 10 min, temperature dependent,
Na+ independent, saturable (Michaelis-Menten constant (Km), 86 +/- 25 microM; maximal velocity (Vmax), 109 +/- 33 nmol (mg
protein)-1 h-1) and reduced by 80-90% by a 50-fold excess concentration of L-glutamate and L-homocysteic acid, but not L-aspartate.
These transport properties resemble those described for system xc-, which exchanges cystine for intracellular glutamate. 3.
Acivicin, a known inhibitor of gamma-GT, decreased gamma-GT activity from 2.58 +/- 0.96 to 0.97 +/- 0.11 mU (mg protein)-1
and decreased the initial rates of L-cystine and L-glutamine uptake by 41-55%. Anthglutin (1-gamma-L-glutamyl-2-(2-carboxyphenylhyl)hydrazine),
a structurally different inhibitor of gamma-GT, also caused a concentration-dependent (0.01-1 mM) decrease in gamma-GT activity
and L-cystine uptake. 4. Neither acivicin nor anthglutin inhibited the uptake of L-glutamate, a poor substrate for gamma-GT.
5. In the presence of a 500-fold excess concentration of glutamate, which should abolish entry of cystine via system xc-,
the remaining fraction of cystine transport was inhibited by 50% by acivicin, suggesting that transport is, in part, dependent
on the activity of gamma-GT. 6. Cystine transport was also 60-80% inhibited by a series of gamma-glutamyl amino acids (5 mM)
including gamma-glutamyl-glutamate, gamma-glutamyl-glutamine and gamma-glutamyl-glycine. alpha-Dipeptides inhibited cystine
transport by only 6-22%. 7. These findings demonstrate that in human pancreatic duct PaTu 8902 cells, cystine uptake is mediated
by system xc- (50-60%) and the gamma-glutamyl cycle. Our results provide the first evidence linking gamma-GT with cystine
transport in human epithelial cells and are of relevance in view of the importance of cystine as a sulphur amino acid source
for GSH synthesis in cells exposed to oxidative stress.</description><identifier>ISSN: 0022-3751</identifier><identifier>EISSN: 1469-7793</identifier><identifier>DOI: 10.1113/jphysiol.1995.sp020721</identifier><identifier>PMID: 7658371</identifier><language>eng</language><publisher>England: The Physiological Society</publisher><subject>Adenosine Triphosphate - metabolism ; Amino Acids - metabolism ; Cell Line ; Cystine - metabolism ; gamma-Glutamyltransferase - antagonists & inhibitors ; gamma-Glutamyltransferase - physiology ; Glutamates - pharmacology ; Humans ; Isoxazoles - pharmacology ; Kinetics ; Microscopy, Electron ; Microscopy, Electron, Scanning ; Pancreatic Ducts - cytology ; Pancreatic Ducts - metabolism ; Pancreatic Ducts - ultrastructure ; Pyrrolidonecarboxylic Acid - pharmacology</subject><ispartof>The Journal of physiology, 1995-05, Vol.485 (Pt 1), p.167-177</ispartof><rights>1995 The Physiological Society</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4187-3ed48fae563055f6020127a4821257a25d9121dc145e8c7d2cf34c922c93299b3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1157981/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1157981/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,315,728,781,785,886,1418,27928,27929,45578,45579,53795,53797</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7658371$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>J H Sweiry</creatorcontrib><creatorcontrib>J Sastre</creatorcontrib><creatorcontrib>J Viña</creatorcontrib><creatorcontrib>H P Elsässer</creatorcontrib><creatorcontrib>G E Mann</creatorcontrib><title>A role for gamma-glutamyl transpeptidase and the amino acid transport system xc- in cystine transport by a human pancreatic duct cell line</title><title>The Journal of physiology</title><addtitle>J Physiol</addtitle><description>1. The roles of the gamma-glutamyl cycle and the anionic amino acid transport system xc- in mediating L-cystine uptake were
investigated in cultured human pancreatic duct PaTu 8902 cells. This cell line exhibits morphological features of normal pancreatic
duct cells and expresses gamma-glutamyl transpeptidase (gamma-GT, EC 2.3.2.2), an enzyme involved in the metabolism and regulation
of intracellular glutathione (GSH). 2. Uptake of L-cystine (10 microM) was linear for up to 10 min, temperature dependent,
Na+ independent, saturable (Michaelis-Menten constant (Km), 86 +/- 25 microM; maximal velocity (Vmax), 109 +/- 33 nmol (mg
protein)-1 h-1) and reduced by 80-90% by a 50-fold excess concentration of L-glutamate and L-homocysteic acid, but not L-aspartate.
These transport properties resemble those described for system xc-, which exchanges cystine for intracellular glutamate. 3.
Acivicin, a known inhibitor of gamma-GT, decreased gamma-GT activity from 2.58 +/- 0.96 to 0.97 +/- 0.11 mU (mg protein)-1
and decreased the initial rates of L-cystine and L-glutamine uptake by 41-55%. Anthglutin (1-gamma-L-glutamyl-2-(2-carboxyphenylhyl)hydrazine),
a structurally different inhibitor of gamma-GT, also caused a concentration-dependent (0.01-1 mM) decrease in gamma-GT activity
and L-cystine uptake. 4. Neither acivicin nor anthglutin inhibited the uptake of L-glutamate, a poor substrate for gamma-GT.
5. In the presence of a 500-fold excess concentration of glutamate, which should abolish entry of cystine via system xc-,
the remaining fraction of cystine transport was inhibited by 50% by acivicin, suggesting that transport is, in part, dependent
on the activity of gamma-GT. 6. Cystine transport was also 60-80% inhibited by a series of gamma-glutamyl amino acids (5 mM)
including gamma-glutamyl-glutamate, gamma-glutamyl-glutamine and gamma-glutamyl-glycine. alpha-Dipeptides inhibited cystine
transport by only 6-22%. 7. These findings demonstrate that in human pancreatic duct PaTu 8902 cells, cystine uptake is mediated
by system xc- (50-60%) and the gamma-glutamyl cycle. Our results provide the first evidence linking gamma-GT with cystine
transport in human epithelial cells and are of relevance in view of the importance of cystine as a sulphur amino acid source
for GSH synthesis in cells exposed to oxidative stress.</description><subject>Adenosine Triphosphate - metabolism</subject><subject>Amino Acids - metabolism</subject><subject>Cell Line</subject><subject>Cystine - metabolism</subject><subject>gamma-Glutamyltransferase - antagonists & inhibitors</subject><subject>gamma-Glutamyltransferase - physiology</subject><subject>Glutamates - pharmacology</subject><subject>Humans</subject><subject>Isoxazoles - pharmacology</subject><subject>Kinetics</subject><subject>Microscopy, Electron</subject><subject>Microscopy, Electron, Scanning</subject><subject>Pancreatic Ducts - cytology</subject><subject>Pancreatic Ducts - metabolism</subject><subject>Pancreatic Ducts - ultrastructure</subject><subject>Pyrrolidonecarboxylic Acid - pharmacology</subject><issn>0022-3751</issn><issn>1469-7793</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1995</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkc9u1DAQxiMEKkvhEUA-cUDKkrHj2L4gtRV_VYkeytnyOs7GlRMH26HkFfrUdbTbqtw4jezvm9_M6CuKd1BtAYB8vJn6JVrvtiAE3capwhXD8KzYQN2IkjFBnhebqsK4JIzCy-JVjDdVBaQS4qQ4YQ3lhMGmuDtDwTuDOh_QXg2DKvduTmpYHEpBjXEyU7KtigapsUWpz3Wwo0dK2_bo8CGhuMRkBvRXl8iOSOeXHc0Tfbcghfp5UCOa1KiDUclq1M46IW2cQy7bXxcvOuWieXOsp8WvL5-vL76Vlz-_fr84uyx1DZyVxLQ175ShDako7Zp8OGCmao4BU6YwbQVgaDXU1HDNWqw7UmuBsRYEC7Ejp8WnA3ead4NptRnznk5OwQ4qLNIrK_9VRtvLvf8jASgTHDLg_REQ_O_ZxCQHG9cz1Gj8HCVjNQfOcTY2B6MOPsZguschUMk1RfmQolxTlA8p5sa3T1d8bDvGlvXzg35rnVn-kyqvf1ytHzWnAA3LkA8HSG_3_a0NRh7aotfWpEVmn7xKEuRqvgd9RMK9</recordid><startdate>19950515</startdate><enddate>19950515</enddate><creator>J H Sweiry</creator><creator>J Sastre</creator><creator>J Viña</creator><creator>H P Elsässer</creator><creator>G E Mann</creator><general>The Physiological Society</general><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><scope>5PM</scope></search><sort><creationdate>19950515</creationdate><title>A role for gamma-glutamyl transpeptidase and the amino acid transport system xc- in cystine transport by a human pancreatic duct cell line</title><author>J H Sweiry ; J Sastre ; J Viña ; H P Elsässer ; G E Mann</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4187-3ed48fae563055f6020127a4821257a25d9121dc145e8c7d2cf34c922c93299b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1995</creationdate><topic>Adenosine Triphosphate - metabolism</topic><topic>Amino Acids - metabolism</topic><topic>Cell Line</topic><topic>Cystine - metabolism</topic><topic>gamma-Glutamyltransferase - antagonists & inhibitors</topic><topic>gamma-Glutamyltransferase - physiology</topic><topic>Glutamates - pharmacology</topic><topic>Humans</topic><topic>Isoxazoles - pharmacology</topic><topic>Kinetics</topic><topic>Microscopy, Electron</topic><topic>Microscopy, Electron, Scanning</topic><topic>Pancreatic Ducts - cytology</topic><topic>Pancreatic Ducts - metabolism</topic><topic>Pancreatic Ducts - ultrastructure</topic><topic>Pyrrolidonecarboxylic Acid - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>J H Sweiry</creatorcontrib><creatorcontrib>J Sastre</creatorcontrib><creatorcontrib>J Viña</creatorcontrib><creatorcontrib>H P Elsässer</creatorcontrib><creatorcontrib>G E Mann</creatorcontrib><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><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of physiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>J H Sweiry</au><au>J Sastre</au><au>J Viña</au><au>H P Elsässer</au><au>G E Mann</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A role for gamma-glutamyl transpeptidase and the amino acid transport system xc- in cystine transport by a human pancreatic duct cell line</atitle><jtitle>The Journal of physiology</jtitle><addtitle>J Physiol</addtitle><date>1995-05-15</date><risdate>1995</risdate><volume>485</volume><issue>Pt 1</issue><spage>167</spage><epage>177</epage><pages>167-177</pages><issn>0022-3751</issn><eissn>1469-7793</eissn><abstract>1. The roles of the gamma-glutamyl cycle and the anionic amino acid transport system xc- in mediating L-cystine uptake were
investigated in cultured human pancreatic duct PaTu 8902 cells. This cell line exhibits morphological features of normal pancreatic
duct cells and expresses gamma-glutamyl transpeptidase (gamma-GT, EC 2.3.2.2), an enzyme involved in the metabolism and regulation
of intracellular glutathione (GSH). 2. Uptake of L-cystine (10 microM) was linear for up to 10 min, temperature dependent,
Na+ independent, saturable (Michaelis-Menten constant (Km), 86 +/- 25 microM; maximal velocity (Vmax), 109 +/- 33 nmol (mg
protein)-1 h-1) and reduced by 80-90% by a 50-fold excess concentration of L-glutamate and L-homocysteic acid, but not L-aspartate.
These transport properties resemble those described for system xc-, which exchanges cystine for intracellular glutamate. 3.
Acivicin, a known inhibitor of gamma-GT, decreased gamma-GT activity from 2.58 +/- 0.96 to 0.97 +/- 0.11 mU (mg protein)-1
and decreased the initial rates of L-cystine and L-glutamine uptake by 41-55%. Anthglutin (1-gamma-L-glutamyl-2-(2-carboxyphenylhyl)hydrazine),
a structurally different inhibitor of gamma-GT, also caused a concentration-dependent (0.01-1 mM) decrease in gamma-GT activity
and L-cystine uptake. 4. Neither acivicin nor anthglutin inhibited the uptake of L-glutamate, a poor substrate for gamma-GT.
5. In the presence of a 500-fold excess concentration of glutamate, which should abolish entry of cystine via system xc-,
the remaining fraction of cystine transport was inhibited by 50% by acivicin, suggesting that transport is, in part, dependent
on the activity of gamma-GT. 6. Cystine transport was also 60-80% inhibited by a series of gamma-glutamyl amino acids (5 mM)
including gamma-glutamyl-glutamate, gamma-glutamyl-glutamine and gamma-glutamyl-glycine. alpha-Dipeptides inhibited cystine
transport by only 6-22%. 7. These findings demonstrate that in human pancreatic duct PaTu 8902 cells, cystine uptake is mediated
by system xc- (50-60%) and the gamma-glutamyl cycle. Our results provide the first evidence linking gamma-GT with cystine
transport in human epithelial cells and are of relevance in view of the importance of cystine as a sulphur amino acid source
for GSH synthesis in cells exposed to oxidative stress.</abstract><cop>England</cop><pub>The Physiological Society</pub><pmid>7658371</pmid><doi>10.1113/jphysiol.1995.sp020721</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Access via Wiley Online Library; IngentaConnect Free/Open Access Journals; PubMed Central; Alma/SFX Local Collection |
| subjects | Adenosine Triphosphate - metabolism Amino Acids - metabolism Cell Line Cystine - metabolism gamma-Glutamyltransferase - antagonists & inhibitors gamma-Glutamyltransferase - physiology Glutamates - pharmacology Humans Isoxazoles - pharmacology Kinetics Microscopy, Electron Microscopy, Electron, Scanning Pancreatic Ducts - cytology Pancreatic Ducts - metabolism Pancreatic Ducts - ultrastructure Pyrrolidonecarboxylic Acid - pharmacology |
| title | A role for gamma-glutamyl transpeptidase and the amino acid transport system xc- in cystine transport by a human pancreatic duct cell line |
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