Conopeptides

Details Conopeptides Conopeptides

1998-08-19

2001-10-23

Low, Christopher S. F. (Department: 1653)

Chemistry: natural resins or derivatives; peptides or proteins;

Peptides of 3 to 100 amino acid residues

25 or more amino acid residues in defined sequence

C530S326000, C530S333000, C530S344000, C514S008100, C514S012200, C514S013800

Reexamination Certificate

active

06307014

ABSTRACT:

BACKGROUND OF THE INVENTION
The invention relates to conopeptides.
Cone snails (Conus) are widely dispersed marine gastropods that prey on fish, marine worms, and mollusks. Cone snails are venomous predators that elaborate peptide toxins in their venom ducts; the venom flows from the venom ducts through hollow stinging radular teeth, into the prey of the snails.
Several of these peptide toxins, generically referred to as conopeptides, have been characterized. The conotoxins are small peptides with a high density of disulfide bonds; cysteine residues can represent up to 50% of the amino acids in some conotoxins. Conotoxins target nicotinic acetylcholine receptors, sodium channels, and calcium channels, and thus behave as potent receptor antagonists.
Another group of peptide toxins includes the conantokins. These peptides, in contrast to other conotoxins, generally contain no cysteine residues. Conantokins are characterized by the presence of one or more &ggr;-carboxyglutamic acid (Gla) residue. Conantokin-G, isolated from the cone snail
Conus geographus,
has 17 residues, 5 of which are Gla residues; conantokin-T, isolated from
Conus tulipa
, has 21 residues, 4 of which are Gla residues. Many of the conantokins have N-methyl-D-aspartate (NMDA) antagonist activity.
Still other conopeptides have some of the characteristics of both the conotoxins and the conantokins. For example, the conopeptide Ctx TxVIIA, isolated from
Conus textile,
contains both Gla residues and cysteine residues.
SUMMARY OF THE INVENTION
The invention features substantially pure Gla-containing peptides. The peptides have the amino acid sequences:
(a) Asp Val Pro X
1
Ile Val Leu X
2
Phe Met Cys Pro Val Ile Cys Gly Asn Gly Phe Gly X
3
Glu Tyr Cys Asn Cys Thr (SEQ ID NO:1), where each of X
1
, X
2
, and X
3
is independently selected from Glu and Gla, provided that at least one of X
1
, X
2
, and X
3
is Gla;
(b) Ser Cys Asp Ser X
4
Phe Ser Ser X
5
Phe Cys X
6
Arg Pro X
7
X
8
Ser Cys Ser Cys Ser Thr His Thr Cys Cys His Trp Ala Arg Arg Asp Gln Cys Met Lys Pro Gln Arg Cys Ile Ser Ala Gln Lys Gly Asn (SEQ ID NO:2), where each of X
4
, X
5
, X
6
, X
7
, and X
8
is independently selected from Glu and Gla, provided that at least one of X
4
, X
5
, X
6
, X
7
, and X
8
is Gla;
(c) Arg X
9
X
10
Cys Cys Ser Asp Pro Arg Cys Asn Ser Ser His Pro X
11
Leu Cys Gly (SEQ ID NO:3), where X
9
is Pro or OHP and each of X
10
and X
11
is independently selected from Glu and Gla, provided that at least one of X
10
and X
11
is Gla;
(d) Gly Cys Asn Asn Ser Cys Gln X
12
His Ser Asp Cys X
13
Ser His Cys Ile Cys Thr Phe Arg Gly Cys Gly Ala Val Asn (SEQ ID NO:4), where each of X
12
and X
3
is independently selected from Glu and Gla, provided that at least one of X
12
and X
13
is Gla;
(e) Cys Ile Pro Gla Gly Ser Ser Cys Ser Ser Ser Gly Ser Cys Cys His Lys Ser Cys Cys Arg Trp Thr Cys Asn Gln Pro Cys Leu (SEQ ID NO:5);
(f) Gly Met X
14
Gly X
15
Cys Lys Asp Gly Leu Thr Thr Cys Leu Ala X
16
Ser X
17
Cys Cys Ser X
18
Asp Cys X
19
Gly Ser Cys Thr Met X
20
(SEQ ID NO:6), where each of X
14
and X
20
is independently selected from Trp and BrW, each of X
15
, X
17
, X
18
, and X
19
is independently selected from Glu and Gla, provided that at least one of X
15
, X
17
, X
18
, and X
19
is Gla, and X
16
is Pro or OHP;
(g) X
21
Cys Cys X
22
Asp Gly X
23
Cys Cys Thr Ala Ala X
24
(SEQ ID NO:7), where each of X
21
and X
22
is independently selected from Glu and Gla, provided that at least one of X
21
and X
22
is Gla, X
23
is Trp or BrW, and X
24
is Pro or OHP;
(h) Ser Cys Ser Asp Asp Trp Gln Tyr Cys Gla Ser X
25
Thr Asp Cys Cys Ser X
26
Asp Cys Asp Val Val Cys Ser (SEQ ID NO:8), where X
25
is Pro or OHP and X
26
is Trp or BrW;
(i) Ala X
27
Cys His X
28
Cys X
29
Phe Gla Tyr (SEQ ID NO:9), where X
27
is Trp or BrW and each of X
28
and X
29
is independently selected from Pro and OHP;
(j) Asn Cys Ser Asp Asp Trp Gln Tyr Cys Gla Ser X
30
Ser Asp Cys Cys Ser X
31
Asp Cys Asp Val Val Cys Ser (SEQ ID NO:10), where X
30
is Pro or OHP and X
31
is Trp or BrW; or
(k) Leu Cys X
32
Asp Tyr Thr X
33
X
34
Cys Ser His Ala His X
35
Cys Cys Ser X
36
Asn Cys Tyr Asn Gly His Cys Thr Gly (SEQ ID NO:11), where each of X
32
and X
34
is independently selected from Pro and OHP, and each of X
33
and X
35
is independently selected from Glu and Gla, provided that at least one of X
33
and X
35
is Gla, and X
36
is Trp or BrW.
“Gla” represents a &ggr;-carboxyglutamic acid residue; “OHP” represents a hydroxyproline residue; and BrW represents a bromotryptophan residue.
One or more of the residues of these peptides may be glycosylated. The termini of the peptides may be modified as well; for example, the C terminus may be amidated.
By “substantially pure” is meant that a conopeptide of the invention has been separated from components which naturally accompany it, or which are generated during its preparation or extraction. Preferably the peptide is at least 90%, more preferably at least 95%, and most preferably at least 99%, by weight, free from the other peptides and molecules with which it is naturally associated. The purity of the peptides can be measured by any appropriate method, for example, column chromatography, polyacrylamide gel electrophoresis, or HPLC analysis.
The peptides of the invention have a variety of applications. They can be used as medicinal agents, for example, as pain medications. They can also be used in veterinary applications; the peptides may be used to temporarily stun, yet not kill, fish and other marine life. Such an application is useful when studying marine life forms.


REFERENCES:
Chandler et al., “Polyamine-like actions of peptides derived from conatokin-G, an N-methyl-D-aspartate (NMDA) antagonist.”J. Biol. Chem., 268:17173-17178 (1993).
Cruz et al., “A preliminary study of Conus venom protein.”Veliger, 18:302-308 (1976).
Cruz et al., “Conus peptides: Phylogenetic range of biological activity.”Biol. Bull., 183:159-164 (1992).
Czerwiec et al., “Vitamin K-dependent carboxylase: Comparison of the bovine y-carboxylase with the y-carboxylase from the marine snail.”Blood 88, (Suppl. 1):523a (1996).
Fainzilber et al., “Mollusc-specific toxins from the venom of Conus textile neovicarius.”Eur. J. Biochem., 588-595 (1991).
Haack et al., “Conantokin-T: a y-carboxyglutamate containing peptide with N-methyl-D-aspartate antagonist activity”J. Biol. Chem., 265:6025-6029 (1989).
McIntosh et al., “y-carboxyglutamate in a neuroactive toxin,”J. Biol. Chem., 259:14343-14346 (1984).
Mena et al., “Conantokin-G: A novel peptide antagonist to the N-methyl-D-aspartic (NMDA) receptor.”Neurosci. Lett., 118:241-244 (1990).
Nakamura et al., “Mass spectrometric-based revision of the structure of a cysteine-rich peptide toxin with y-carboxyglutamic acid, TxVIIA, from the sea snail, Conus textile,”Protein Sci., 5:524-530 (1996).
Olivera et al., “Peptide neurotoxins from fish-hunting cone snails.”Science, 230:1338-1343 (1985).
Olivera et al., “Diversity of Conus neuropeptides.” Science, 249:257-263 (1990).
Olivera et al., “Conotoxins.”J. Biol. Chem., 33:22067-22070 (1991).
Rigby et al., “Three dimensional structure of a y-carboxyglutamic acid-containing conotoxin, conantokin-G, from the marine cone snail Conus geographus: The metal-free conformer.”Biochemistry36:6906-6914 (1997).
Rigby et al., “The role of y-carboxyglutamic acid in the calcium-induced structural transition of conantokin-G, a contoxin from the marine cone snail Conus geographus.”Biochemistry36:15677-15684 (1997).
Skjaerbaek et al., “Determination of the solution structures of conantokin-G and conantokin-T by CD and NMR spectroscopy.”J. Biol. Chem., 272:2291-2299 (1997).
Woodward et al., “Constant and hypervariable regions in conotoxin propeptides.”EMBOJ., 9:1051-1020 (1990).
Zhou et al., “Synthetic analogues of conantokin G: NMDA antagonists acting through a novel polyamine-coupled site.”J. Neurochem., 66:620-628 (1996).

Affiliated with

Also associated with

Rating

Conopeptides does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Conopeptides, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Conopeptides will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2561364