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Abstract

Grant Number:	1R43HL073827-01
PI Name:	LEDIZET, MICHEL E.
PI Email:	l2dx@aol.com
PI Title:
Project Title:	A Novel Family of Anticoagulants from Ixodes scapularis

Abstract: DESCRIPTION (provided by applicant): The overall objective of this proposal is to evaluate the anticoagulant properties of a family of novel tick salivary proteins Ixodes scapularis, the black-legged tick, circumvents host hemostatic mechanisms as it engorges on vertebrate hosts. This tick's saliva contains potent anticoagulants, including a Factor Xa inhibitor. Purified salivary proteins with anticoagulant activity were subjected to N-terminal sequencing. A corresponding tick salivary cDNA was cloned that encoded a 96 kDa protein, designated Salp9A. This protein is homologous to Salp14, which we previously identified by immunoscreening a tick salivary gland cDNA expression library. Additional protein and cDNA sequencing indicate that I scapularis salivary glands express at least ten different Salp14-related proteins. We expressed and purified recombinant Salp14, and found that it inhibits coagulation and Factor Xa. These data suggest that we have discovered a new family of anticoagulants. In Phase I experiments we will obtain full-length cDNAs encoding selected Salp14 family members. Recombinant proteins will be expressed from these cDNAs. The recombinant proteins will be compared to partially purified salivary anticoagulants in anticoagulant and protease inhibition assays. These studies will relate structure and function, and provide a rational basis for selecting anticoagulant molecules for Phase II animal studies.

Thesaurus Terms:
Ixodes, anticoagulant, blood coagulation, drug discovery /isolation, endopeptidase, protein structure function
coagulation factor X, complementary DNA, gene expression, information system, inhibitor /antagonist, posttranslational modification, protease inhibitor, recombinant protein, salivary gland, structural model
Drosophilidae, Escherichia coli, computer assisted sequence analysis

Institution:	L2 DIAGNOSTICS, LLC
	BOX 8175
	NEW HAVEN, CT 94904
Fiscal Year:	2003
Department:
Project Start:	22-AUG-2003
Project End:	31-AUG-2004
ICD:	NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
IRG:	ZRG1

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