title: Conformational modulation mediated by polyglutamine expansion in CAG repeat expansion disease-associated proteins.
abstract: We have previously reported TR-FRET based immunoassays to detect a conformational change imparted on huntingtin protein by the polyglutamine expansion, which we confirmed using biophysical methodologies. Using these immunoassays, we now report that polyglutamine expansion influences the conformational properties of other polyglutamine disease proteins, exemplified by the androgen receptor (associated with spinal bulbar muscular atrophy) and TATA binding protein (associated with spinocerebellar ataxia 17). Using artificial constructs bearing short or long polyglutamine expansions or a multimerized, unrelated epitope (mimicking the increase in anti-polyglutamine antibody epitopes present in polyglutamine repeats of increasing length) we confirmed that the conformational TR-FRET based immunoassay detects an intrinsic conformational property of polyglutamine repeats. The TR-FRET based conformational immunoassay may represent a rapid, scalable tool to identify modulators of polyglutamine-mediated conformational change in different proteins associated with CAG triplet repeat disorders.
authors: IRBM Promidis, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy; IRBM Science Park, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy.
authors: Margherita
authors: M
authors: Verani
authors: IRBM Science Park, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy.
authors: Maria
authors: M
authors: Bustamante
authors: IRBM Science Park, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy.
authors: Paola
authors: P
authors: Martufi
authors: IRBM Science Park, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy.
authors: Manuel
authors: M
authors: Daldin
authors: IRBM Science Park, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy.
authors: Cristina
authors: C
authors: Cariulo
authors: IRBM Promidis, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy; IRBM Science Park, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy.
authors: Lucia
authors: L
authors: Azzollini
authors: IRBM Promidis, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy; IRBM Science Park, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy.
authors: Valentina
authors: V
authors: Fodale
authors: IRBM Science Park, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy.
authors: Francesca
authors: F
authors: Puglisi
authors: IRBM Promidis, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy.
authors: Andreas
authors: A
authors: Weiss
authors: CHDI Management/CHDI Foundation, Los Angeles, CA, 90045, USA.
authors: Douglas
authors: D
authors: Macdonald
authors: IRBM Promidis, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy; IRBM Science Park, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy.
authors: Lara
authors: L
authors: Petricca
authors: IRBM Promidis, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy; IRBM Science Park, Via Pontina km 30.600, 00071, Pomezia, Rome, Italy. Electronic address: a.caricasole@irbm.it.
authors: Andrea
authors: A
authors: Caricasole
mesh_terms: D002457:Cell Extracts; D004194:Disease / Q000235:genetics*; D031541:Fluorescence Resonance Energy Transfer; D057809:HEK293 Cells; D006801:Humans; D007118:Immunoassay; D008968:Molecular Conformation; D010455:Peptides / Q000378:metabolism*; D014162:Transfection; D019680:Trinucleotide Repeat Expansion / Q000235:genetics*
chemical_list: D002457:Cell Extracts; D010455:Peptides; C097188:polyglutamine
keywords: CAG repeat; Conformation; Huntingtin; Neurodegeneration; Polyglutamine; TR–FRET assay