| Project/Area Number |
09044331
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Field |
General pharmacology
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| Research Institution | Kyorin University |
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Principal Investigator |
KANAI Yoshikatdu Kyorin University, school of Medicine, Associato Professor, 医学部, 助教授 (60204533)
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| Co-Investigator(Kenkyū-buntansha) |
HOSOYAMADA Makoto Kyorin University, school of Medicine, Assistant, 医学部, 助手 (00291659)
SEKINE Takashi Kyorin University, school of Medicine, Assistant, 医学部, 助手 (50255402)
ENDOU Hitoshoi Kyorin University, school of Medicine, Professor, 医学部, 教授 (20101115)
TATE Naoko (UTSUNOMIYA Naoko) Kyorin University, school of Medicine, Assistant, 医学部, 助手 (00201955)
TROTTI David ハーバード大学, 医学部, 助手
HEDIGER Matt ハーバード大学, 医学部, 準教授
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥9,100,000 (Direct Cost: ¥9,100,000)
Fiscal Year 1998: ¥4,400,000 (Direct Cost: ¥4,400,000)
Fiscal Year 1997: ¥4,700,000 (Direct Cost: ¥4,700,000)
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| Keywords | Transporter / Membrane transport / amino acids / oxidative stress / cystine / cysteine / Central nervous system / Molewlar Pharmawlogy / 神経細胞 / クローニング / グリオーマ |
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| Research Abstract |
In the central nervous system, transporters for amino acids such as glutamate and cysteine or cystine are present on the plasma membrane of neurons and gliai cells and blood-brain barrier to protect neurons against excitotoxicity or oxidative stress. In the present study. Ye have performed cDNA cloning to elucidate the molecular nature of transporters for cysteine or cystine.
First, we performed expression cloning in which mRNA from rat C6 glioma cells were co-expressed with 4F2 heavy chain (4F2hc). At the end, we isolated a cDNA encoding. a 512 amino acid protein designated LAT1 (L-type amino acid transporter 1). In addition, in the process of the cloning of LAT1, we established the protocol for the "co-expression cloning" using Xenopus laevis oocytes. When expressed in Xenopus oocytes, LAT1 required 4F2hc for its functional expression. The co-expression of LAT1 and 4F2hc induced the function of classically characterized amino acid transport system L which transports large neutral amin
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o acids with NaィイD1+ィエD1-independent manner. Because LAT1 is expressed in the central nervous system and transports L-dopa, phenylalanine, tyrosine and tryptophan, it is suggested that LAT1 plays roles, not only in cellular nutrition but also In monoamine metabolism in brain.
Based on the cDNA sequence of LAT1, we searched the EST (expressed sequence tag) data base for LAT1-related proteins. We have identified 5 related sequences (EST1〜5). We cloned full length cDNAs, and found that EST3 (designated LAT2) is a transporter which transports all the neutral α-amino acids including cysteine. LAT2 also required 4F2hc for its functional expression. LAT2 is expressed in brain, small intestine, kidney and placenta, suggesting its involvement in the permeation at blood-tissue barrier and transepithelial transport.
Second, we isolated a cDNA for cyctine transporter BAT1 with the properties of classical system bィイD10ィエD1,+. BAT1 associated with rBAT structurally related to 4F2hc to be functional. BAT1 was expressed in neurons in brain and seemed to be important for the protection of neurons from oxidative stress. Less
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