IDENTIFICATION AND MOLECULAR CHARACTERISATION OF PROTEINS, EXPRESSED IN THE IXODES RICINUS SALIVARY GLANDS
The invention relates to a new polynucleotide which encodes a polypeptide expressed in the salivary glands of ticks, more particularly the Ixodes ricinus arthropod tick, during the slow-feeding phase of the blood meal have, said polynucleotide and related polypeptide may be used in different constructions and for different applications which are also included in the present invention.
1 . A method for providing a desired benefit to an individual comprising administering a sufficient amount of an isolated polypeptide having at least 75% sequence identity to the amino acid sequence of SEQ. ID. NO: 36 or a sufficient amount of an isolated polypeptide having at least 75% sequence identity to the amino acid sequence of SEQ. ID. NO: 36 fused to at least one additional heterologous amino acid sequence to said individual to provide said desired benefit, wherein said desired benefit is selected from the group consisting of (i) inhibition of a plasma contact factor, (ii) ameliorating a condition for which inhibition of the intrinsic coagulation pathway maybe beneficial, (iii) reducing the risk of bleeding while inhibiting clot formation, and (iv) reducing the risk of thrombosis.
2 . The method of claim 1 , wherein the desired benefit is reducing the risk of arterial thrombosis.
3 . The method of claim 1 wherein there is administered a sufficient amount of an isolated polypeptide comprising at least 85% sequence identity to the amino acid sequence of SEQ. ID. NO: 36.
4 . The method of claim 1 , wherein said desired benefit is inhibition of activation of a plasma contact factor and wherein the activation of said plasma contact factor is generated through contact between a polyanionic surface and factor XII.
5 . The method of claim 1 wherein said desired benefit is inhibition of the activation of a plasma contact factor and wherein said inhibition of the activation of a plasma contact factor has an effect selected from the group consisting of inhibition of clot formation without interfering with clotting equilibrium and inhibition of clot formation with reduced risk of interfering with bleeding time.
6 . The method of claim 1 , wherein said inhibition of a plasma contact factor has an effect selected from reduction in the risk of pulmonary embolism, reduction in the risk of cerebral thrombosis or ischemia, or reduction in the risk of deep vein thrombosis.
7 . The method of claim 1 , wherein the polypeptide inhibits the activation of factor XIIa, factor XIa and prekallikrein.
8 . The method of claim 7 , wherein the polypeptide inhibits the activation of prekallikrein into kallikrein by factor XIIa, the activation of factor XI into factor XIa by factor XIIa and the activation of factor XII into factor XIIa by factor XIa.
9 . The method of claim 1 , wherein the polypeptide does not have a specific interaction with prekallikrein, HMWK, factor XII, factor XI, factor IX, factor IXa, factor X, factor Xa, thrombin, factor VIIa, t-PA and plasminogen at a level similarly high to that of its interaction with factor XIIa, factor XIa and kallikrein.
10 . The method of claim 1 , wherein the polypeptide does not have a specific interaction with prekallikrein, HMWK, factor XII, factor XI, factor IX, factor IXa, factor X, factor Xa, thrombin, factor VIIa, t-PA and plasminogen at a level similarly high to that of its interaction with factor XIIa, factor XIa, plasmin and kallikrein.
11 . The method of claim 1 , wherein said condition is a condition for which inhibition of the intrinsic coagulation pathway is beneficial and is a condition for which inhibition of factor VIIa and factor Xa is less beneficial.
12 . The method of claim 1 , wherein said individual is a mammal.
13 . The method of claim 1 , wherein the polypeptide which is administered to said individual is an isolated polypeptide consisting of the amino acid sequence of SEQ ID NO: 36 or an isolated polypeptide consisting of the amino acid sequence of SEQ ID NO: 36 fused to at least one additional heterologous amino acid sequence.
14 . The method of claim 1 , wherein at least one amino acid in said polypeptide has a substitution group.
15 . The method of claim 1 , wherein at least one amino acid in said polypeptide has a substitution group selected from the group consisting of amide, acetyl, phosphoryl, and glycosyl groups.
16 . The method of claim 1 , wherein said heterologous amino acid sequence comprises multiple histidine residues.
17 . The method of claim 1 , wherein the polypeptide is present in a pharmaceutical composition further comprising an adequate pharmaceutical carrier.
18 . The method of claim 1 , wherein the polypeptide comprises at least 90% percent sequence identity with the amino acid sequence of SEQ ID NO: 36.
19 . The method of claim 1 , wherein the polypeptide comprises at least 95% sequence identity with SEQ ID NO: 36 .
20 . The method of claim 1 , wherein the polypeptide is present in a pharmaceutical composition further comprising an adequate pharmaceutical carrier.
21 . The method of claim 1 , wherein the polypeptide comprises at least 90% sequence identity with SEQ ID NO: 36 .
22 . The method of claim 12 , wherein the polypeptide comprises at least 95% sequence identity with SEQ ID NO: 36 .
23 . The method of claim 12 , wherein the mammal is a human.