{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T00:30:44Z","timestamp":1791937844368,"version":"4.3.4"},"reference-count":50,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2012,11,22]],"date-time":"2012-11-22T00:00:00Z","timestamp":1353542400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["The FEBS Journal"],"published-print":{"date-parts":[[2013,1]]},"abstract":"<jats:p>Vertebrate photoreceptors contain a unique tetraspanin protein known as \u2018retinal degeneration slow\u2019 (<jats:styled-content style=\"fixed-case\">RDS<\/jats:styled-content>). Mutations in the <jats:italic><jats:styled-content style=\"fixed-case\">RDS<\/jats:styled-content><\/jats:italic> gene have been identified in a variety of human retinal degenerative diseases, and more than 70% of these mutations are located in the second intra\u2010discal (<jats:styled-content style=\"fixed-case\">D<\/jats:styled-content>2) loop, highlighting the importance of this region. Here we examined the conformational and thermal stability properties of the <jats:styled-content style=\"fixed-case\">D<\/jats:styled-content>2 loop of <jats:styled-content style=\"fixed-case\">RDS<\/jats:styled-content>, as well as interactions with <jats:styled-content style=\"fixed-case\">ROM<\/jats:styled-content>\u20131, a non\u2010glycosylated homolog of <jats:styled-content style=\"fixed-case\">RDS<\/jats:styled-content>. The <jats:styled-content style=\"fixed-case\">RDS D<\/jats:styled-content>2 loop was expressed in <jats:italic>Escherichia\u00a0coli<\/jats:italic> as a fusion protein with maltose binding protein (<jats:styled-content style=\"fixed-case\">MBP<\/jats:styled-content>). The fusion protein, referred to as <jats:styled-content style=\"fixed-case\">MBP<\/jats:styled-content>\u2013<jats:styled-content style=\"fixed-case\">D<\/jats:styled-content>2, was purified to homogeneity. Circular dichroism spectroscopy showed that the wild\u2010type (<jats:styled-content style=\"fixed-case\">WT<\/jats:styled-content>) <jats:styled-content style=\"fixed-case\">D<\/jats:styled-content>2 loop consists of approximately 21% \u03b1\u2013helix, approximately 20% \u03b2\u2013sheet and approximately 59% random coil. <jats:styled-content style=\"fixed-case\">D<\/jats:styled-content>2 loop fusion proteins carrying disease\u2010causing mutations in <jats:styled-content style=\"fixed-case\">RDS<\/jats:styled-content> (e.g. <jats:styled-content style=\"fixed-case\">R<\/jats:styled-content>172<jats:styled-content style=\"fixed-case\">W<\/jats:styled-content>,<jats:styled-content style=\"fixed-case\"> C<\/jats:styled-content>214<jats:styled-content style=\"fixed-case\">S<\/jats:styled-content>,<jats:styled-content style=\"fixed-case\"> N<\/jats:styled-content>244<jats:styled-content style=\"fixed-case\">H<\/jats:styled-content>\/<jats:styled-content style=\"fixed-case\">K<\/jats:styled-content>) were also examined, and conformational changes were observed (compared to wild\u2010type <jats:styled-content style=\"fixed-case\">D<\/jats:styled-content>2). In particular, the <jats:styled-content style=\"fixed-case\">C<\/jats:styled-content>150S, <jats:styled-content style=\"fixed-case\">C<\/jats:styled-content>214<jats:styled-content style=\"fixed-case\">S<\/jats:styled-content> and <jats:styled-content style=\"fixed-case\">N<\/jats:styled-content>244<jats:styled-content style=\"fixed-case\">H<\/jats:styled-content> proteins showed significant reductions in \u03b1\u2013helicity. However, the thermal stability of the mutants was unchanged compared to wild\u2010type, and all the mutants were capable of interacting with <jats:styled-content style=\"fixed-case\">ROM<\/jats:styled-content>\u20131, indicating that this functional aspect of the isolated <jats:styled-content style=\"fixed-case\">D<\/jats:styled-content>2 loop remained intact in the mutants despite the observed conformational changes. An I\u2013<jats:styled-content style=\"fixed-case\">TASSER<\/jats:styled-content> model of the <jats:styled-content style=\"fixed-case\">RDS D<\/jats:styled-content>2 loop predicted a structure consistent with the circular dichroism experiments and the structure of the conserved region of the <jats:styled-content style=\"fixed-case\">D<\/jats:styled-content>2 loop of other tetraspanin family members. These results provide significant insight into the mechanism of <jats:styled-content style=\"fixed-case\">RDS<\/jats:styled-content> complex formation and the disease process underlying <jats:styled-content style=\"fixed-case\">RDS<\/jats:styled-content>\u2010associated retinal degeneration.<\/jats:p>","DOI":"10.1111\/febs.12055","type":"journal-article","created":{"date-parts":[[2012,11,2]],"date-time":"2012-11-02T09:01:37Z","timestamp":1351846897000},"page":"127-138","source":"Crossref","is-referenced-by-count":8,"title":["Structural characterization of the second intra\u2010discal loop of the photoreceptor tetraspanin <scp>RDS<\/scp>"],"prefix":"10.1111","volume":"280","author":[{"given":"Dibyendu","family":"Chakraborty","sequence":"first","affiliation":[{"name":"Department of Cell Biology University of Oklahoma Health Sciences Center Oklahoma City OK USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karla K.","family":"Rodgers","sequence":"additional","affiliation":[{"name":"Department of Biochemistry &amp; Molecular Biology University of Oklahoma Health Sciences Center Oklahoma City OK USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shannon M.","family":"Conley","sequence":"additional","affiliation":[{"name":"Department of Cell Biology University of Oklahoma Health Sciences Center Oklahoma City OK USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muna I.","family":"Naash","sequence":"additional","affiliation":[{"name":"Department of Cell Biology University of Oklahoma Health Sciences Center Oklahoma City OK 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