{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,26]],"date-time":"2025-10-26T14:18:06Z","timestamp":1761488286474},"reference-count":40,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2009,7,20]],"date-time":"2009-07-20T00:00:00Z","timestamp":1248048000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Proteins"],"published-print":{"date-parts":[[2010,2]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Some neurodegenerative diseases such as Alzheimer disease (AD) and Parkinson disease are caused by protein misfolding. In AD, amyloid \u03b2\u2010peptide (A\u03b2) is thought to be a toxic agent by self\u2010assembling into a variety of aggregates involving soluble oligomeric intermediates and amyloid fibrils. Here, we have designed several green fluorescent protein (GFP) variants that contain pseudo\u2010A\u03b2 \u03b2\u2010sheet surfaces and evaluated their abilities to bind to A\u03b2 and inhibit A\u03b2 oligomerization. Two GFP variants P13H and AP93Q bound tightly to A\u03b2, <jats:italic>K<\/jats:italic><jats:sub>d<\/jats:sub> = 260 n<jats:italic>M<\/jats:italic> and <jats:italic>K<\/jats:italic><jats:sub>d<\/jats:sub> = 420 n<jats:italic>M<\/jats:italic>, respectively. Moreover, P13H and AP93Q were capable of efficiently suppressing the generation of toxic A\u03b2 oligomers as shown by a cell viability assay. By combining the P13H and AP93Q mutations, a super variant SFAB4 comprising four strands of A\u03b2\u2010derived sequences was designed and bound more tightly to A\u03b2 (<jats:italic>K<\/jats:italic><jats:sub>d<\/jats:sub> = 100 n<jats:italic>M<\/jats:italic>) than those having only two pseudo\u2010A\u03b2 strands. The SFAB4 protein preferentially recognized the soluble oligomeric intermediates of A\u03b2 more than both unstructured monomer and mature amyloid fibrils. Thus, the design strategy for embedding pseudo\u2010A\u03b2 \u03b2\u2010sheet structures onto a protein surface arranged in the \u03b2\u2010barrel structure is useful to construct molecules capable of binding tightly to A\u03b2 and inhibiting its aggregation. This strategy may provide implication for the diagnostic and therapeutic development in the treatment of AD. Proteins 2010. \u00a9 2009 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/prot.22546","type":"journal-article","created":{"date-parts":[[2009,7,21]],"date-time":"2009-07-21T14:09:44Z","timestamp":1248185384000},"page":"336-347","source":"Crossref","is-referenced-by-count":16,"title":["Rational design of amyloid \u03b2 peptide\u2013binding proteins: Pseudo\u2010A\u03b2 \u03b2\u2010sheet surface presented in green fluorescent protein binds tightly and preferentially to structured 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