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Structure‐based design of inhibitory peptide for End Binding proteins
Author(s) -
Saqib Uzma,
Geyer Melissa,
Minshall Richard,
Malik Asrar,
Komarova Yulia
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.1122.6
Subject(s) - microbiology and biotechnology , peptide , chemistry , histamine , paracellular transport , nitric oxide , calcium in biology , endothelium , receptor , biochemistry , biology , pharmacology , endocrinology , permeability (electromagnetism) , organic chemistry , membrane
The Microtubule (MT) cytoskeleton plays an important role in regulating paracellular permeability of the endothelium in the context of inflammation; however, the molecular mechanism involved in MT‐dependent increase in permeability is not yet fully understood. We have shown that End Binding (EB) protein 3, which binds to and regulates MT dynamics in the endothelium, interacts with ER associated inositol 1,4,5‐trisphosphate (IP3) receptor type 3 and facilitates thrombin‐induced release of calcium from ER stores. Based on the crystal structure of the EB interacting domain and using the computational alanine scan approach, we designed a 7‐amino acid synthetic inhibitory peptide that mimics interaction between EB3 and IP3R3. We have shown that treatment of human pulmonary artery endothelial cells with this peptide blocked the interaction between IP3R3 and EB3 and negatively regulated thrombin‐ and histamine‐induced increase in intracellular calcium. Consistent with this finding, EB inhibitory peptide attenuated histamine‐induced activation of endothelial nitric oxide synthase (eNOS), as determined by NO production but had no effect on basal level of NO. In an animal model of inflammation, EB peptide prevented histamine‐induces vasodilation. Our data suggests that EB proteins can be used as a potential target for development of novel therapies to prevent leaky vessel syndrome during inflammation. F unding source : RO1 HL103922 to Y ulia K omarova