Regulation of amino acid/carnitine transporter B⁰,⁺ (ATB⁰,⁺) in astrocytes by protein kinase C: independent effects on raft and non-raft transporter subpopulations

J. Neurochem. (2010) 115, 1386-1397. ABSTRACT: Neutral and basic amino acid transporter B⁰,⁺ belongs to a Na,Cl-dependent superfamily of proteins transporting neurotransmitters, amino acids and osmolytes, known to be regulated by protein kinase C (PKC). The present study demonstrates an increased ph...

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Veröffentlicht in:Journal of neurochemistry 2010-12, Vol.115 (6), p.1386-1397
Hauptverfasser: Samluk, Łukasz, Czeredys, Magdalena, Nałęcz, Katarzyna A
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Czeredys, Magdalena
Nałęcz, Katarzyna A
description J. Neurochem. (2010) 115, 1386-1397. ABSTRACT: Neutral and basic amino acid transporter B⁰,⁺ belongs to a Na,Cl-dependent superfamily of proteins transporting neurotransmitters, amino acids and osmolytes, known to be regulated by protein kinase C (PKC). The present study demonstrates an increased phosphorylation of B⁰,⁺ on serine moiety after treatment of rat astrocytes with phorbol 12-myristate 13-acetate, a process correlated with an augmented activity of l-leucine transport and an enhanced presence of the transporter at the cell surface. After solubilization with Triton X-100 and sucrose gradient centrifugation, B⁰,⁺ was detected in non-raft as well as in detergent-resistant raft fractions under control conditions, while phorbol 12-myristate 13-acetate treatment resulted in a complete disappearance of the transporter from the raft fraction. B⁰,⁺ was observed to interact with caveolin-1 and flotillin-1 (reggie-2) proteins, the markers of detergent-resistant microdomains of plasma membrane. As verified in immunocytochemistry and immunoprecipitation experiments, modification of PKC activity did not affect these interactions. It is proposed that PKC reveals different effects on raft and non-raft subpopulations of B⁰,⁺. Phorbol ester treatment results in trafficking of the transporter from the intracellular pool to non-raft microdomains and increased activity, while B⁰,⁺ present in raft microdomains undergoes either internalization or is transferred laterally to non-raft domains.
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Neurochem. (2010) 115, 1386-1397. ABSTRACT: Neutral and basic amino acid transporter B⁰,⁺ belongs to a Na,Cl-dependent superfamily of proteins transporting neurotransmitters, amino acids and osmolytes, known to be regulated by protein kinase C (PKC). The present study demonstrates an increased phosphorylation of B⁰,⁺ on serine moiety after treatment of rat astrocytes with phorbol 12-myristate 13-acetate, a process correlated with an augmented activity of l-leucine transport and an enhanced presence of the transporter at the cell surface. After solubilization with Triton X-100 and sucrose gradient centrifugation, B⁰,⁺ was detected in non-raft as well as in detergent-resistant raft fractions under control conditions, while phorbol 12-myristate 13-acetate treatment resulted in a complete disappearance of the transporter from the raft fraction. 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Neuroglia ; Kinases ; L-leucine ; Membrane and intracellular transports ; membrane rafts ; Molecular and cellular biology ; Neurochemistry ; Neurotransmitters ; phorbol 12-myristate 13-acetate ; Phorbol esters ; Phosphorylation ; Plasma membranes ; Protein kinase C ; Protein transport ; Serine ; Solubilization ; Sucrose ; transporter translocation ; Vertebrates: nervous system and sense organs</subject><ispartof>Journal of neurochemistry, 2010-12, Vol.115 (6), p.1386-1397</ispartof><rights>2010 The Authors. 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Neurochem. (2010) 115, 1386-1397. ABSTRACT: Neutral and basic amino acid transporter B⁰,⁺ belongs to a Na,Cl-dependent superfamily of proteins transporting neurotransmitters, amino acids and osmolytes, known to be regulated by protein kinase C (PKC). The present study demonstrates an increased phosphorylation of B⁰,⁺ on serine moiety after treatment of rat astrocytes with phorbol 12-myristate 13-acetate, a process correlated with an augmented activity of l-leucine transport and an enhanced presence of the transporter at the cell surface. After solubilization with Triton X-100 and sucrose gradient centrifugation, B⁰,⁺ was detected in non-raft as well as in detergent-resistant raft fractions under control conditions, while phorbol 12-myristate 13-acetate treatment resulted in a complete disappearance of the transporter from the raft fraction. B⁰,⁺ was observed to interact with caveolin-1 and flotillin-1 (reggie-2) proteins, the markers of detergent-resistant microdomains of plasma membrane. As verified in immunocytochemistry and immunoprecipitation experiments, modification of PKC activity did not affect these interactions. It is proposed that PKC reveals different effects on raft and non-raft subpopulations of B⁰,⁺. Phorbol ester treatment results in trafficking of the transporter from the intracellular pool to non-raft microdomains and increased activity, while B⁰,⁺ present in raft microdomains undergoes either internalization or is transferred laterally to non-raft domains.</description><subject>Amino acids</subject><subject>Astrocytes</subject><subject>B0,+ transporter</subject><subject>Biological and medical sciences</subject><subject>Carnitine</subject><subject>caveolin-1</subject><subject>Cell physiology</subject><subject>Cell surface</subject><subject>Cellular biology</subject><subject>Centrifugation</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Immunocytochemistry</subject><subject>Immunoprecipitation</subject><subject>Isolated neuron and nerve. Neuroglia</subject><subject>Kinases</subject><subject>L-leucine</subject><subject>Membrane and intracellular transports</subject><subject>membrane rafts</subject><subject>Molecular and cellular biology</subject><subject>Neurochemistry</subject><subject>Neurotransmitters</subject><subject>phorbol 12-myristate 13-acetate</subject><subject>Phorbol esters</subject><subject>Phosphorylation</subject><subject>Plasma membranes</subject><subject>Protein kinase C</subject><subject>Protein transport</subject><subject>Serine</subject><subject>Solubilization</subject><subject>Sucrose</subject><subject>transporter translocation</subject><subject>Vertebrates: nervous system and sense organs</subject><issn>0022-3042</issn><issn>1471-4159</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><recordid>eNpdkc1u1DAQxyMEEkvhGbCQEFQi27GdxNlKHNoVn6pAgvZsTRy78pK1g-0V3WOfhwtnHqdPgtNdKoQP9njmN1_6FwWhMKf5HK3mtBK0rGi9mDPIXhBQwfzqXjG7C9wvZgCMlRwq9rB4FOMKgDZVQ2fFzy_6cjNgst4RbwiurfMEle2PFAZnk3WapIAujj4kHcjpzfWvVzfXv8nLk_O_9iGxjmBMwatt0pF0WzIGn3T2frMOoybL44z0etT5coloY7RKkeSeAU0i6HrivCtvP_92i5tu9ON-vvi4eGBwiPrJ_j0oLt6-OV--L88-v_uwPDkrDQcGZYXUNEphh5wL2guOtQJoBTN13dWM9V3f9AsBC9FRVYmO99BTYYC3FTRNZ_hB8WJXN2_xfaNjkmsblR4GdNpvomzrRogWaJvJZ_-RK78JLg8nW1oznpEmQ8_3EEaFg8n7KRvlGOwaw1ayjEDb8My93nE_7KC3d3EKchJaruSkp5z0lJPQ8lZoeSU_flpOVs5_uss36CVehtzj4msmOdBFHiQTfwC-Nqz6</recordid><startdate>201012</startdate><enddate>201012</enddate><creator>Samluk, Łukasz</creator><creator>Czeredys, Magdalena</creator><creator>Nałęcz, Katarzyna A</creator><general>Blackwell Publishing Ltd</general><general>Wiley-Blackwell</general><scope>FBQ</scope><scope>IQODW</scope><scope>7QR</scope><scope>7TK</scope><scope>7U7</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope></search><sort><creationdate>201012</creationdate><title>Regulation of amino acid/carnitine transporter B⁰,⁺ (ATB⁰,⁺) in astrocytes by protein kinase C: independent effects on raft and non-raft transporter subpopulations</title><author>Samluk, Łukasz ; Czeredys, Magdalena ; Nałęcz, Katarzyna A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-f3020-4a1f6ccaba3371d73a5c00872f55b522dbd6d97097b1c47b3d0d17f0384066bf3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Amino acids</topic><topic>Astrocytes</topic><topic>B0,+ transporter</topic><topic>Biological and medical sciences</topic><topic>Carnitine</topic><topic>caveolin-1</topic><topic>Cell physiology</topic><topic>Cell surface</topic><topic>Cellular biology</topic><topic>Centrifugation</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Immunocytochemistry</topic><topic>Immunoprecipitation</topic><topic>Isolated neuron and nerve. Neuroglia</topic><topic>Kinases</topic><topic>L-leucine</topic><topic>Membrane and intracellular transports</topic><topic>membrane rafts</topic><topic>Molecular and cellular biology</topic><topic>Neurochemistry</topic><topic>Neurotransmitters</topic><topic>phorbol 12-myristate 13-acetate</topic><topic>Phorbol esters</topic><topic>Phosphorylation</topic><topic>Plasma membranes</topic><topic>Protein kinase C</topic><topic>Protein transport</topic><topic>Serine</topic><topic>Solubilization</topic><topic>Sucrose</topic><topic>transporter translocation</topic><topic>Vertebrates: nervous system and sense organs</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Samluk, Łukasz</creatorcontrib><creatorcontrib>Czeredys, Magdalena</creatorcontrib><creatorcontrib>Nałęcz, Katarzyna A</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Chemoreception Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Toxicology 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>Biotechnology and BioEngineering Abstracts</collection><jtitle>Journal of neurochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Samluk, Łukasz</au><au>Czeredys, Magdalena</au><au>Nałęcz, Katarzyna A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Regulation of amino acid/carnitine transporter B⁰,⁺ (ATB⁰,⁺) in astrocytes by protein kinase C: independent effects on raft and non-raft transporter subpopulations</atitle><jtitle>Journal of neurochemistry</jtitle><date>2010-12</date><risdate>2010</risdate><volume>115</volume><issue>6</issue><spage>1386</spage><epage>1397</epage><pages>1386-1397</pages><issn>0022-3042</issn><eissn>1471-4159</eissn><coden>JONRA9</coden><abstract>J. Neurochem. (2010) 115, 1386-1397. ABSTRACT: Neutral and basic amino acid transporter B⁰,⁺ belongs to a Na,Cl-dependent superfamily of proteins transporting neurotransmitters, amino acids and osmolytes, known to be regulated by protein kinase C (PKC). The present study demonstrates an increased phosphorylation of B⁰,⁺ on serine moiety after treatment of rat astrocytes with phorbol 12-myristate 13-acetate, a process correlated with an augmented activity of l-leucine transport and an enhanced presence of the transporter at the cell surface. After solubilization with Triton X-100 and sucrose gradient centrifugation, B⁰,⁺ was detected in non-raft as well as in detergent-resistant raft fractions under control conditions, while phorbol 12-myristate 13-acetate treatment resulted in a complete disappearance of the transporter from the raft fraction. B⁰,⁺ was observed to interact with caveolin-1 and flotillin-1 (reggie-2) proteins, the markers of detergent-resistant microdomains of plasma membrane. As verified in immunocytochemistry and immunoprecipitation experiments, modification of PKC activity did not affect these interactions. It is proposed that PKC reveals different effects on raft and non-raft subpopulations of B⁰,⁺. Phorbol ester treatment results in trafficking of the transporter from the intracellular pool to non-raft microdomains and increased activity, while B⁰,⁺ present in raft microdomains undergoes either internalization or is transferred laterally to non-raft domains.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><doi>10.1111/j.1471-4159.2010.07040.x</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record>
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subjects Amino acids
Astrocytes
B0,+ transporter
Biological and medical sciences
Carnitine
caveolin-1
Cell physiology
Cell surface
Cellular biology
Centrifugation
Fundamental and applied biological sciences. Psychology
Immunocytochemistry
Immunoprecipitation
Isolated neuron and nerve. Neuroglia
Kinases
L-leucine
Membrane and intracellular transports
membrane rafts
Molecular and cellular biology
Neurochemistry
Neurotransmitters
phorbol 12-myristate 13-acetate
Phorbol esters
Phosphorylation
Plasma membranes
Protein kinase C
Protein transport
Serine
Solubilization
Sucrose
transporter translocation
Vertebrates: nervous system and sense organs
title Regulation of amino acid/carnitine transporter B⁰,⁺ (ATB⁰,⁺) in astrocytes by protein kinase C: independent effects on raft and non-raft transporter subpopulations
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