Amino acid transporters and uses

• US 5,658,782 A
• Filed: 10/20/1993
• Issued: 08/19/1997
• Est. Priority Date: 10/20/1993
• Status: Expired due to Fees

• Current Assignee: Oregon State University (Oregon University System)
• Original Assignee: Oregon State University (Oregon University System)
• Inventors: Amara, Susan G., Arriza, Jeffrey L.
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): HOBBS, LISA JOE
• Application Number:
  US08/140,729
• Time in Patent Office: 1,399 Days
• Field of Search: 435/240.2, 435/320.1, 530/350, 536/24.31, 536/23.5
• US Class Current: 435/365
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 2799/023 where the vector is derived from a poxvirus

Abstract

The present invention relates to novel mammalian amino acid transporter proteins and the genes that encode such proteins. The invention is directed toward the isolation, characterization and pharmacological use of the human amino acid transporter proteins EAAT1, EAAT2, EAAT3 and ASCT1. The invention specifically provides isolated complementary DNA copies of mRNA corresponding to each of these transporter genes. Also provided are recombinant expression constructs capable of expressing each of the amino acid transporter genes of the invention in cultures of transformed prokaryotic and eukaryotic cells, as well as such cultures of transformed cells that synthesize the human amino acid transporter proteins encoded therein. The invention also provides methods for screening in vitro compounds having transport-modulating properties using preparations of transporter proteins from such cultures of cells transformed with recombinant expression constructs.

Claims

1. An isolated nucleic acid comprising a nucleotide sequence encoding a human excitatory amino acid transporter selected from the group consisting of the glutamate transporter EAAT2.

2. An isolated nucleic acid according to claim 1 wherein the nucleotide sequence of the nucleic acid comprises the sequence of the human excitatory amino acid transporter EAAT2 (SEQ ID No.:

6).

4. A nucleic acid hybridization probe for the detection of mammalian excitatory amino acid transporter expression comprising the nucleotide sequence of claim 2.

3. A homogeneous composition of a mammalian excitatory amino acid transporter having a molecular weight of about 62.1 kilodaltons and an amino acid sequence comprising the amino acid sequence of the human excitatory amino acid transporter EAAT2 (SEQ ID No.:

7).

5. A recombinant expression construct comprising a nucleic acid having a nucleotide sequence encoding a mammalian excitatory amino acid transporter, wherein the nucleotide sequence comprises the sequence of the human EAAT2 excitatory amino acid transporter (SEQ ID No.:

6), and wherein the construct is capable of expressing the EAAT2 excitatory amino acid transporter in a transformed culture of eukaryotic or prokaryotic cells.

6. A cell culture transformed with the recombinant expression construct of claim 5, wherein the transformed cell culture expresses the EAAT2 excitatory amino acid transporter.