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Vol. 16, Issue 4, 1725-1734, April 2005
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* Department of Pathology, Emory University, Atlanta, GA 30322;
Bioorganic Chemistry, Genentech, South San Francisco, CA 94080;
School of Pharmacy, King's College London, Strand, London WC2R 2LS, United Kingdom; and
Cardiff University, School of Pharmacy, Cardiff, Wales CF10 3XF
Submitted June 9, 2004;
Revised January 6, 2005;
Accepted January 10, 2005
Monitoring Editor: Daniel Goodenough
Occludin is a tetraspan integral membrane protein in epithelial and endothelial tight junction (TJ) structures that is projected to have two extracellular loops. We have used peptides emulating central regions of human occludin's first and second loops, termed O-A:101121 and O-B:210228, respectively, to examine potential molecular interactions between these two regions of occludin and other TJ proteins. A superficial biophysical assessment of A:101121 and O-B:210228 showed them to have dissimilar solution conformation characteristics. Although O-A:101121 failed to strongly interact with protein components of the human epithelial intestinal cell line T84, O-B:210228 selectively associated with occludin, claudin-one and the junctional adhesion molecule (JAM)-A. Further, the presence of O-B:210228, but not O-A:101121, impeded the recovery of functional TJ structures. A scrambled peptide sequences of O-B:210228 failed to influence TJ assembly. These studies demonstrate distinct properties for these two extracellular segments of the occludin protein and provide an improved understanding of how specific domains of occludin may interact with proteins present at TJ structures.
Abbreviations used: TJ, tight junction; JAM, junctional adhesion molecule; TER, trans-epithelial electric resistance; CD, circular dichroism.
Address correspondence to: Asma Nusrat (anusrat{at}emory.edu).
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