WO2001004293A1 - Polypeptides et polynucleotides associes a un transporteur de glycine - Google Patents

Polypeptides et polynucleotides associes a un transporteur de glycine Download PDF

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Publication number
WO2001004293A1
WO2001004293A1 PCT/EP2000/006254 EP0006254W WO0104293A1 WO 2001004293 A1 WO2001004293 A1 WO 2001004293A1 EP 0006254 W EP0006254 W EP 0006254W WO 0104293 A1 WO0104293 A1 WO 0104293A1
Authority
WO
WIPO (PCT)
Prior art keywords
polypeptide
sequence
polynucleotide
seq
isolated
Prior art date
Application number
PCT/EP2000/006254
Other languages
English (en)
Inventor
Hugh Jonathan Herdon
Joanne Rachel Evans
Original Assignee
Smithkline Beecham P.L.C.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Smithkline Beecham P.L.C. filed Critical Smithkline Beecham P.L.C.
Priority to EP00956163A priority Critical patent/EP1194553A1/fr
Publication of WO2001004293A1 publication Critical patent/WO2001004293A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals

Definitions

  • polypeptide sequence of SEQ ID NO:2 (e) the polypeptide sequence of SEQ ID NO:2; and (f) an isolated polypeptide having or comprising a polypeptide sequence that has an Identity Index of 0.95, 0.96, 0.97, 0.98, or 0.99 compared to the polypeptide sequence of SEQ ID NO:2; (g) fragments and variants of such polypeptides in (a) to (f); with the proviso that the isolated polypeptide does not have the ammo acid sequence of SEQ ID NO:4.
  • an isolated polynucleotide having or comprising a polynucleotide sequence encoding a polypeptide sequence that has an Identity Index of 0.95, 0.96, 0.97, 0.98, or 0.99 compared to the polypeptide sequence of SEQ ID NO:2; and polynucleotides that are fragments and variants of the above mentioned polynucleotides or that are complementary to above mentioned polynucleotides, over the entire length thereof; with the proviso that the isolated polynucleotide does not have the sequence of SEQ ID NO:3.
  • the present invention provides polynucleotides that are RNA transcripts of the
  • Polynucleotides of the present invention may be obtained using standard cloning and screening techniques from a cDNA library de ⁇ ved from mRNA in cells of human brain and hippocampus (see for instance, Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor
  • Preferred probes and primers will generally comprise at least 15 nucleotides, preferably, at least 30 nucleotides and may have at least 50, if not at least 100 nucleotides. Particularly preferred probes will have between 30 and 50 nucleotides. Particularly preferred primers will have between 20 and 25 nucleotides.
  • Such agonists or antagonists so-identified may be natural or modified substrates, ligands, receptors, enzymes, etc., as the case may be, of the polypeptide; a structural or functional mimetic thereof (see Coligan et al., Current Protocols in Immunology l(2):Chapter 5 (1991)) or a small molecule.
  • Such small molecules preferably have a molecular weight below 2,000 daltons, more preferably between 300 and 1,000 daltons, and most preferably between 400 and 700 daltons. It is preferred that these small molecules are organic molecules.
  • a variant and reference polypeptide may differ in amino acid sequence by one or more substitutions, insertions, deletions in any combination.
  • a substituted or inserted amino acid residue may or may not be one encoded by the genetic code. Typical conservative substitutions include Gly, Ala; Val, He, Leu; Asp, Glu; Asn, Gin; Ser, Thr; Lys, Arg; and Phe and Tyr.
  • a variant of a polynucleotide or polypeptide may be naturally occurring such as an allele, or it may be a variant that is not known to occur naturally.
  • Non-naturally occurring variants of polynucleotides and polypeptides may be made by mutagenesis techniques or by direct synthesis. Also included as variants are polypeptides having one or more post-translational modifications, for instance glycosylation, phosphorylation, methylation, ADP ribosylation and the like. Embodiments include methylation of the N-terminal amino acid, phosphorylations of serines and threonines and modification of C-terminal glycines. "Allele” refers to one of two or more alternative forms of a gene occuring at a given locus in the genome.
  • Suspensions of cells transiently transfected as described in Example 1 are pipetted into polylysine-coated 96 well plates (50,000 cells per well). 4-24 hours later, the wells are washed with standard Krebs/HEPES buffer (KHB) at 23°C. After addition of further KHB to the wells, the plates incubated at 23°C for 10 min. KHB containing appropriate concentrations of a potential inhibitor is then added to the wells, together with 0.5 ⁇ M [ ⁇ H]-glycine. After incubation at 23°C for 2-60 min, the wells are washed with ice-cold KHB. The cells are then solubilised, and the amount of radioactivity present determined using a scintillation counter.
  • KHB standard Krebs/HEPES buffer

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biochemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Zoology (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Toxicology (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention concerne GC42 qui est un polypeptide semblable à un transporteur de glycine. L'invention concerne des polypeptides et des polynucleotides GC42, ainsi que des méthodes de production de ces polypeptides par des techniques de recombinaison. L'invention concerne également des méthodes d'utilisation des polypeptides et des polynucleotides GC42 dans des méthodes diagnostiques.
PCT/EP2000/006254 1999-07-09 2000-07-04 Polypeptides et polynucleotides associes a un transporteur de glycine WO2001004293A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP00956163A EP1194553A1 (fr) 1999-07-09 2000-07-04 Polypeptides et polynucleotides associes a un transporteur de glycine

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9916195.2 1999-07-09
GBGB9916195.2A GB9916195D0 (en) 1999-07-09 1999-07-09 Novel compounds

Publications (1)

Publication Number Publication Date
WO2001004293A1 true WO2001004293A1 (fr) 2001-01-18

Family

ID=10857011

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2000/006254 WO2001004293A1 (fr) 1999-07-09 2000-07-04 Polypeptides et polynucleotides associes a un transporteur de glycine

Country Status (3)

Country Link
EP (1) EP1194553A1 (fr)
GB (1) GB9916195D0 (fr)
WO (1) WO2001004293A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993010228A1 (fr) * 1991-11-12 1993-05-27 Synaptic Pharmaceutical Corporation And codant un transporteur de glycine et ses emplois
WO1997045446A1 (fr) * 1996-05-31 1997-12-04 Allelix Neuroscience Inc. Transporteur de glycine et cellules transfectees et leurs emplois
WO1998046619A1 (fr) * 1997-04-11 1998-10-22 Allelix Neuroscience Inc. Transporteur de la glycine

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993010228A1 (fr) * 1991-11-12 1993-05-27 Synaptic Pharmaceutical Corporation And codant un transporteur de glycine et ses emplois
WO1997045446A1 (fr) * 1996-05-31 1997-12-04 Allelix Neuroscience Inc. Transporteur de glycine et cellules transfectees et leurs emplois
WO1998046619A1 (fr) * 1997-04-11 1998-10-22 Allelix Neuroscience Inc. Transporteur de la glycine

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
KIM K -M ET AL: "CLONING OF THE HUMAN GLYCINE TRANSPORTER TYPE 1: MOLECULAR AND PHARMACOLOGICAL CHARACTERIZATION OF NOVEL ISOFORM VARIANTS AND CHROMOSOMAL LOCALIZATION OF THE GENE IN THE HUMAN AND MOUSE GENOMES", MOLECULAR PHARMACOLOGY,US,BALTIMORE, MD, vol. 45, 1994, pages 608 - 617, XP002911694, ISSN: 0026-895X *

Also Published As

Publication number Publication date
GB9916195D0 (en) 1999-09-15
EP1194553A1 (fr) 2002-04-10

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