CA2405450A1 - Dermacentor variabilis gaba-gated chloride channels - Google Patents

Dermacentor variabilis gaba-gated chloride channels Download PDF

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Publication number
CA2405450A1
CA2405450A1 CA002405450A CA2405450A CA2405450A1 CA 2405450 A1 CA2405450 A1 CA 2405450A1 CA 002405450 A CA002405450 A CA 002405450A CA 2405450 A CA2405450 A CA 2405450A CA 2405450 A1 CA2405450 A1 CA 2405450A1
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CA
Canada
Prior art keywords
seq
polypeptide
group
gated chloride
unique
Prior art date
Legal status (The legal status 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 status listed.)
Granted
Application number
CA002405450A
Other languages
French (fr)
Other versions
CA2405450C (en
Inventor
Yingcong Zheng
Doris Cully
Steven Ludmerer
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Boehringer Ingelheim Animal Health USA Inc
Original Assignee
Individual
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 Individual filed Critical Individual
Publication of CA2405450A1 publication Critical patent/CA2405450A1/en
Application granted granted Critical
Publication of CA2405450C publication Critical patent/CA2405450C/en
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

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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
    • C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
    • C07K14/43513—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae
    • C07K14/43527—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae from ticks
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • A61P33/14—Ectoparasiticides, e.g. scabicides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Abstract

The present invention features Dermacentor variabilis GABA-gated chloride channel polypeptides and nucleic acids, and uses of such polypeptides and nucleic acids. D. variabilis is a widely distributed tick associated with different diseases. A preferred use of the present invention is to obtain compounds for preventing or treating a tick infestation.

Claims (33)

1. A purified polypeptide comprising a unique amino acid region of SEQ. ID. NO. 1, SEQ. ID. NO. 2, or SEQ. ID. NO. 3 that is at least 9 contiguous amino acids in length, wherein said unique region is not present in SEQ. ID.
NO. 7.
2. The polypeptide of claim 1, wherein said unique region comprises an amino acid sequence selected from the group consisting of:
QILNAFFTRG (SEQ. ID. NO. 9);
MTVGAEVAERIWVP (SEQ. ID. NO. 10);
RWSDGDTSVRIAK (SEQ. ID. NO. 11);
TALLEYAAVGYLG (SEQ. ID. NO. 12);
RCAAASSNEPSSEPLLASPEVSIVKT (SEQ. ID. NO. 13);
QPREAPPTGFT (SEQ. ID. NO. 14);
MGRRGADQCCPGLQGSCQVC (SEQ. ID. NO. 15);
MEVRLKMVDPKGFSKSS (SEQ. ID. NO. 16);
HISDVLPDDVGDD (SEQ. ID. NO. 17); and HVSDVLPDDVGDD (SEQ. ID. NO. 18).
3. The polypeptide of claim 1, wherein said polypeptide comprises an amino acid sequence selected from the group consisting of SEQ.
ID.
NO. 1, SEQ. ID. NO. 2, and SEQ. ID. NO. 3.
4. The polypeptide of claim 3, wherein said polypeptide consists of an amino acid sequence selected from the group consisting of SEQ. ID. NO.
1, SEQ. ID. NO. 2, and SEQ. ID. NO. 3.
5. The polypeptide of claim 1, wherein said polypeptide consists of an amino acid sequence having a sequence identity of at least 70% with SEQ.
ID.
NO. 1.
6. A purified nucleic acid comprising a nucleotide sequence encoding for the polypeptide of claim 1.
7. A purified nucleic acid comprising a nucleotide sequence encoding for the polypeptide of claim 5.
8. A purified nucleic acid comprising a unique nucleotide sequence region of SEQ. ID. NO. 4, SEQ. ID. NO. 5, SEQ. ID. NO. 6 or the complement thereof, that is at least 18 contiguous nucleotides in length, wherein said unique region is not present in SEQ. ID. NO. 8 or the complement thereof.
9. The purified nucleic acid of claim 8, wherein said unique region comprises a sequence selected from the group consisting of:
CAAACGCAACGTGGACAA (SEQ. ID. NO. 20);
GAGCGACTCTCGTTCCAG (SEQ. ID. NO. 21);
ATCGGCTCCGGAGGAGAG (SEQ. ID. NO. 22);
AAGGTCCTCGGTCACGTCCAAAAA (SEQ. ID. NO. 23);
CTCGGCAAGAGAATCACC (SEQ. ID. NO. 24);
GGTTCCTGTCAAGTTTGT (SEQ. ID. NO. 25);
GGTTCCTGTCGGGTTTGT (SEQ. ID. NO. 26); and CCAACCGGATTTACCATG (SEQ. ID. NO. 27).
10. The purified nucleic acid of claim 8, wherein said nucleic acid comprises a sequence selected from the group consisting of the nucleotide sequence of SEQ. ID. NO. 4, SEQ. ID. NO. 5 and SEQ. ID. NO. 6.
11. The purified nucleic acid of claim 10, wherein said nucleic acid consists of a sequence selected from the group consisting of the nucleotide sequence of SEQ. ID. NO. 4, SEQ. ID. NO. 5 and SEQ. ID. NO. 6.
12. An expression vector comprising a recombinant nucleotide sequence encoding for a unique amino acid region of SEQ. ID. NO. 1, SEQ. ID.
NO.
2, or SEQ. ID. NO. 3 that is at least 9 contiguous amino acids in length, wherein said unique region is not present in SEQ. ID. NO. 7.
13. The expression vector of claim 12, wherein said recombinant nucleotide sequence encodes for a polypeptide comprising a sequence selected from the group consisting of SEQ. ID. NO. 1, SEQ. ID. NO. 2 and SEQ. ID. NO. 3.
14. A recombinant cell comprising the expression vector of claim 12, wherein said recombinant nucleotide sequence is functionally coupled to a promoter recognized by said cell.
15. A recombinant cell comprising the expression vector of claim 13, wherein said recombinant nucleotide sequence is functionally coupled to a promoter recognized by said cell.
16. A recombinant cell made by a process comprising the step of introducing into said cell the expression vector of claim 12, wherein said recombinant nucleotide sequence is functionally coupled to a promoter recognized by said cell.
17. A purified antibody preparation comprising an antibody that binds a polypeptide selected from the group consisting of SEQ. ID. NO. 1, SEQ.
ID.
NO. 2 and SEQ. ID. NO. 3.
18. The antibody of claim 17, wherein said polypeptide consists of the amino acid sequence of SEQ. ID. NO. 19 (LGKRITMRKTRCQQLAKLAEQ
HRQR).
19. A method of preparing a GABA-gated chloride polypeptide comprising the step of incubating a cell comprising a recombinant nucleotide sequence encoding for a unique amino acid region of SEQ. ID. NO. 1, SEQ. ID.
NO.
2, or SEQ. ID. NO. 3 that is at least 9 contiguous amino acids in length, wherein said unique region is not present in SEQ. ID. NO. 7, under conditions wherein said polypeptide is expressed.
20. A method for assaying the binding of a compound to a D.
variabilis GABA-gated chloride channel comprising the steps of:
a) expressing from a recombinant nucleic acid a polypeptide comprising a unique amino acid region of SEQ. ID. NO. 1, SEQ. ID.
NO.
2, or SEQ. ID. NO. 3 that is at least 9 contiguous amino acids in length, wherein said unique region is not present in SEQ. ID. NO. 7;

b) providing to said polypeptide a test preparation comprising one or more test compounds; and c) measuring the ability of said test preparation to bind to said polypeptide.
21. The method of claim 20, wherein said polypeptide consists of 9 contiguous amino acids up to the full-length sequence provided in SEQ. ID.
NOs. 1, 2, or 3.
22. The method of claim 21, wherein said steps (b) and (c) are performed in vitro.
23. The method of claim 21, wherein said steps (a), (b) and (c) are performed using a whole cell.
24. The method of claim 21, wherein said polypeptide is expressed from an expression vector.
25. The method of claim 24, wherein said polypeptide consists of a sequence selected from the group consisting of SEQ. ID. NO. 1, SEQ. ID. NO. 2 and SEQ. ID. NO. 3.
26. A method of measuring GABA-gated chloride channel activity comprising the steps of:
a) contacting a recombinant cell with a compound, wherein said recombinant cell comprises a recombinant nucleotide sequence expressing a functional GABA-gated chloride channel that comprises a unique amino acid region of SEQ. ID. NO. 1, SEQ. ID. NO. 2, or SEQ. ID. NO. 3 that is at least 9 contiguous amino acids in length, provided that said unique region is not present in SEQ. ID. NO. 7; and b) measuring GABA-gated chloride channel activity.
27. The method of claim 26, wherein said functional GABA-gated chloride channel comprises an amino sequence having a sequence identity of at least 70% with SEQ. ID. NO. 1.
28. The method of claim 26, wherein said functional GABA-gated chloride channel comprises a sequence selected from the group consisting of SEQ. ID.
NO. 1, SEQ. ID. NO. 2 and SEQ. ID. NO. 3.
29. The method of claim 26, wherein said functional GABA-gated chloride channel consists of a sequence selected from the group consisting of SEQ.
ID. NO. 1, SEQ. ID. NO. 2 and SEQ. ID. NO. 3.
30. A method of decreasing or preventing a tick infestation comprising the steps of:
a) identifying a compound that modulates a D. variabilis GABA-gated chloride channel activity; and b) using said compound to decrease or prevent a tick infestation.
31. The method of claim 30, wherein said step (a) is performed by (i) contacting a recombinant cell with said compound, wherein said recombinant cell comprises a recombinant nucleotide sequence expressing a functional GABA-gated chloride channel that comprises a unique amino acid region of SEQ. ID. NO. 1, SEQ.
ID. NO. 2, or SEQ. ID. NO. 3 that is at least 9 contiguous amino acids in length, provided that said unique region is not present in SEQ. ID. NO. 7 and (ii) measuring GABA-gated chloride channel activity.
32. The method of claim 31, wherein said functional GABA-gated chloride channel comprises a sequence selected from the group consisting of SEQ. ID.
NO. 1, SEQ. ID. NO. 2, and SEQ. ID. NO. 3.
33. The method of claim 32, wherein said functional GABA-gated chloride channel consists of a sequence selected from the group consisting of SEQ.
ID. NO. 1, SEQ. ID. NO. 2, and SEQ. ID. NO. 3.
CA2405450A 2000-03-31 2001-03-28 Dermacentor variabilis gaba-gated chloride channels Expired - Lifetime CA2405450C (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US19379100P 2000-03-31 2000-03-31
US60/193,791 2000-03-31
PCT/US2001/009955 WO2001074884A1 (en) 2000-03-31 2001-03-28 Dermacentor variabilis gaba-gated chloride channels

Publications (2)

Publication Number Publication Date
CA2405450A1 true CA2405450A1 (en) 2001-10-11
CA2405450C CA2405450C (en) 2011-08-16

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Family Applications (1)

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CA2405450A Expired - Lifetime CA2405450C (en) 2000-03-31 2001-03-28 Dermacentor variabilis gaba-gated chloride channels

Country Status (6)

Country Link
US (1) US7220832B2 (en)
EP (1) EP1294751B1 (en)
JP (1) JP2004500119A (en)
CA (1) CA2405450C (en)
DK (1) DK1294751T3 (en)
WO (1) WO2001074884A1 (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE60123943T2 (en) * 2000-03-31 2007-06-14 Merial Ltd. LIGAND-CONTROLLED ION CHANNELS FROM DERMACENTOR VARIABILIS CODING DNA MOLECULES
US7267964B2 (en) 2000-03-31 2007-09-11 Merial Limited DNA molecules encoding ligand gated ion channels from Dermacentor variabilis

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU650622B2 (en) 1989-07-11 1994-06-30 Gen-Probe Incorporated Nucleic acid sequence amplification methods utilizing a transcription complex
US5852188A (en) 1990-01-11 1998-12-22 Isis Pharmaceuticals, Inc. Oligonucleotides having chiral phosphorus linkages
US5859221A (en) 1990-01-11 1999-01-12 Isis Pharmaceuticals, Inc. 2'-modified oligonucleotides
WO1993007161A1 (en) 1991-10-04 1993-04-15 Cornell Research Foundation, Inc. Molecular cloning and transformation of cyclodiene resistance
US5487976A (en) 1993-10-15 1996-01-30 Cornell Research Foundation, Inc. DNA encoding an insect gamma-aminobutyric acid (GABA) receptor subunit cells expressing it, and pesticide screening methods using such cells
US5527703A (en) 1994-05-25 1996-06-18 Merck & Co., Inc. DNA encoding glutamate gated chloride channels
US5693492A (en) 1995-05-05 1997-12-02 Merck & Co., Inc. DNA encoding glutamate gated chloride channels
US5661035A (en) 1995-06-07 1997-08-26 The Regents Of The University Of California Voltage sensing by fluorescence resonance energy transfer
US5767261A (en) 1995-11-08 1998-06-16 Rhone -Poulenc Ag Company Lepidopteran GABA gated chloride channel and nucleic acids encoding subunits thereof
US6130038A (en) 1996-07-16 2000-10-10 Gen-Probe Incorporated Method for amplifying target nucleic acids using modified primers
US5854002A (en) 1996-12-17 1998-12-29 Rhone Poulenc, Inc. Method of identifying compounds that bind to the insect gaba receptor
US6329516B1 (en) * 1997-04-28 2001-12-11 Fmc Corporation Lepidopteran GABA-gated chloride channels

Also Published As

Publication number Publication date
WO2001074884A1 (en) 2001-10-11
JP2004500119A (en) 2004-01-08
DK1294751T3 (en) 2014-03-24
US20030208041A1 (en) 2003-11-06
US7220832B2 (en) 2007-05-22
EP1294751B1 (en) 2014-03-05
EP1294751A4 (en) 2004-10-13
EP1294751A1 (en) 2003-03-26
CA2405450C (en) 2011-08-16

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