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The Screening of PDZ‐Domain Array to Identify JAM‐B Interacting Proteins
Author(s) -
Samant Ritika,
Naik Ulhas
Publication year - 2009
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.23.1_supplement.877.3
Subject(s) - pdz domain , transmembrane protein , transmembrane domain , chemistry , microbiology and biotechnology , cytoplasm , protein–protein interaction , biology , biochemistry , gene , receptor
Tight junctions (TJs) host complexes that include PDZ domain‐containing proteins. PDZ proteins participate in transmembrane molecule organization and TJ formation. Transmembrane protein Junctional Adhesion Molecule‐A (JAM‐A) is found in TJs and interacts with PDZ proteins AF‐6 and ZO‐1 via its PDZ binding motif. Another JAM family member, JAM‐B displays such a motif but PDZ binding partners are unknown. Because of the similar roles and cytoplasmic regions of JAM‐A and JAM‐B, we hypothesize that JAM‐B also binds to PDZ proteins. To identify these interactions, HA‐tagged JAM‐B will be used to probe a protein microarray spotted with PDZ domains. A cDNA construct encoding a HA‐tagged GST JAM‐B protein was created. E. coli BL21 cells transformed with this construct were induced by IPTG, and recombinant protein expression and solubility was analyzed by SDS‐PAGE. JAM‐B ‐ PDZ domain interactions on the array will be detected by chemiluminescence. This analysis of JAM‐B and PDZ protein interactions will further our understanding of signaling pathways particularly pertaining to TJ formation and regulation. Supported by an HHMI Undergraduate Science Education Grant, Beckman Foundation and the NIH.