Novel Dimer Compounds That Bind α-Synuclein Can Rescue Cell Growth in a Yeast Model Overexpressing α-Synuclein. A Possible Prevention Strategy for Parkinson's Disease.
The misfolding of α-synuclein is a critical event in the death of dopaminergic neurons and the progression of Parkinson's disease. Previously, it was suggested that drugs, which bind to α-synuclein and form a loop structure between the N- and C-termini, tend to be neuroprotective, whereas others, which cause a more compact structure, tend to be neurotoxic. To improve the binding to α-synuclein, eight novel compounds were synthesized from a caffeine scaffold attached to (R,S)-1-aminoindan, (R,S)-nicotine, and metformin, and their binding to α-synuclein determined through nanopore analysis and isothermal titration calorimetry. The ability of the dimers to interact with α-synuclein in a cell system was assayed in a yeast model of PD which expresses an AS-GFP (α-synuclein-Green Fluorescent Protein) construct under the control of a galactose promoter. In 5 mM galactose this yeast strain will not grow and large cytoplasmic foci are observed by fluorescent microscopy. Two of the dimers, C8-6-I and C8-6-N, at a concentration of 0.1 μM allowed the yeast to grow normally in 5 mM galactose and the AS-GFP became localized to the periphery of the cell. Both dimers were superior when compared to the monomeric compounds. The presence of the dimers also caused the disappearance of preformed cytoplasmic foci. Nanopore analysis of C8-6-I and C8-6-N were consistent with simultaneous binding to both the N- and C-terminus of α-synuclein but the binding constants were only 105M-1.
Department of Biochemistry, ‡Drug Discovery and Development Research Group, College of Pharmacy and Nutrition, and §Department of Anatomy and Cell Biology, University of Saskatchewan , 107 Wiggins Road, Saskatoon, Canada S7N 0W0.
Joe
J
Kakish
Department of Biochemistry, ‡Drug Discovery and Development Research Group, College of Pharmacy and Nutrition, and §Department of Anatomy and Cell Biology, University of Saskatchewan , 107 Wiggins Road, Saskatoon, Canada S7N 0W0.
Kevin J H
KJ
Allen
Department of Biochemistry, ‡Drug Discovery and Development Research Group, College of Pharmacy and Nutrition, and §Department of Anatomy and Cell Biology, University of Saskatchewan , 107 Wiggins Road, Saskatoon, Canada S7N 0W0.
Troy A
TA
Harkness
Department of Biochemistry, ‡Drug Discovery and Development Research Group, College of Pharmacy and Nutrition, and §Department of Anatomy and Cell Biology, University of Saskatchewan , 107 Wiggins Road, Saskatoon, Canada S7N 0W0.
Ed S
ES
Krol
Department of Biochemistry, ‡Drug Discovery and Development Research Group, College of Pharmacy and Nutrition, and §Department of Anatomy and Cell Biology, University of Saskatchewan , 107 Wiggins Road, Saskatoon, Canada S7N 0W0.
Jeremy S
JS
Lee
D000978:Antiparkinson Agents / Q000138:chemical synthesis / Q000494:pharmacology*; D002151:Calorimetry; D019281:Dimerization; D004353:Drug Evaluation, Preclinical; D005690:Galactose / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010300:Parkinson Disease / Q000517:prevention & control; D011485:Protein Binding; D015003:Yeasts / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism*
D000978:Antiparkinson Agents; D051844:alpha-Synuclein; D049452:Green Fluorescent Proteins; D005690:Galactose
Parkinson’s disease; druggable yeast; isothermal titration calorimetry; nanopore analysis; neuroprotective drugs; neurotoxic drugs; protein folding; α-synuclein