title: Unaided trifluoroacetic acid pretreatment solubilizes polyglutamine peptides and retains their biophysical properties of aggregation.
abstract: Understanding the biophysical mechanism of polyglutamine (polyGln) aggregation is important to unravel the role of aggregates in the pathology of polyGln repeat disorders. To achieve this, synthetic polyGln peptides are widely used. Their disaggregation and solubilization is essential because it plays a crucial role in reproducing biophysical experimental data under in vitro conditions. Pretreatment with trifluoroacetic acid (TFA) and hexafluoroisopropanol (HFIP) at a 1:1 ratio is currently the method of choice to achieve solubility of polyGln peptides. Here we report that the disaggregation and solubilization of polyGln peptides can be achieved by TFA alone. We tested TFA due to the close similarity of it with HFIP in the nature of H-bond breakage and formation, higher cost, and the problems faced by us in the availability of HFIP. Our results demonstrate that the TFA disaggregated polyGln sequences give similar solubilization yield, aggregation kinetics, thioflavin T (ThT) binding, and structural features in comparison with the TFA/HFIP method. Furthermore, we show by limited validation studies that the proposed TFA method can replace the existing TFA/HFIP disaggregation method of polyGln sequences.
authors.affiliation: Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur 208 016, India.
authors.forename: Gunasekhar
authors.initials: G
authors.lastname: Burra
authors.affiliation: Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur 208 016, India. Electronic address: akthakur@iitk.ac.in.
authors.forename: Ashwani Kumar
authors.initials: AK
authors.lastname: Thakur
mesh_terms: D052160:Benzothiazoles; D007700:Kinetics; D018625:Microscopy, Atomic Force; D046529:Microscopy, Electron, Transmission; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D020005:Propanols / Q000737:chemistry; D011485:Protein Binding; D012995:Solubility; D017550:Spectroscopy, Fourier Transform Infrared; D013844:Thiazoles / Q000737:chemistry / Q000378:metabolism; D014269:Trifluoroacetic Acid / Q000737:chemistry*; D006845:Hydrocarbons, Fluorinated
chemical_list: D052160:Benzothiazoles; D010455:Peptides; D020005:Propanols; D013844:Thiazoles; D014269:Trifluoroacetic Acid; C009462:thioflavin T; C097188:polyglutamine; C001337:hexafluoroisopropanol; D006845:Hydrocarbons, Fluorinated
keywords: Disaggregation; Hexafluoroisopropanol; Huntington's disease; Polyglutamine; Solubilization; Trifluoroacetic acid