title: A hidden aggregation-prone structure in the heart of hypoxia inducible factor prolyl hydroxylase.
abstract: Prolyl hydroxylase domain-containing protein 2 (PHD2), as one of the most important regulators of angiogenesis and metastasis of cancer cells, is a promising target for cancer therapy drug design. Progressive studies imply that abnormality in PHD2 function may be due to misfolding. Therefore, study of the PHD2 unfolding pathway paves the way for a better understanding of the influence of PHD2 mutations and cancer cell metabolites on the protein folding pathway. We study the unfolding of the PHD2 catalytic domain using differential scanning calorimetry (DSC), fluorescence spectroscopy, and discrete molecular dynamics simulations (DMD). Using computational and experimental techniques, we find that PHD2 undergoes four transitions along the thermal unfolding pathway. To illustrate PHD2 unfolding events in atomic detail, we utilize DMD simulations. Analysis of computational results indicates an intermediate species in the PHD2 unfolding pathway that may enhance aggregation propensity, explaining mutation-independent PHD2 malfunction.
authors: Department of Biological Sciences, Institute for Advanced Studies in Basic Sciences (IASBS), Zanjan, Iran.|Institute of Biochemistry and Biophysics (IBB), University of Tehran, Tehran, Iran.
authors: Hamid
authors: H
authors: Hadi-Alijanvand
authors: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, 02139.
authors: Elizabeth A
authors: EA
authors: Proctor
authors: Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, School of Medicine, Chapel Hill, North Carolina, 27599.|Department of Physics and Astronomy, Clemson University, Clemson, South Carolina, 29634.
authors: Feng
authors: F
authors: Ding
authors: Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, School of Medicine, Chapel Hill, North Carolina, 27599.|Curriculum in Bioinformatics and Computational Biology, University of North Carolina at Chapel Hill, School of Medicine, Chapel Hill, North Carolina, 27599.|Program in Molecular and Cellular Biophysics, University of North Carolina at Chapel Hill, School of Medicine, Chapel Hill, North Carolina, 27599.
authors: Nikolay V
authors: NV
authors: Dokholyan
authors: Institute of Biochemistry and Biophysics (IBB), University of Tehran, Tehran, Iran.|Center of Excellence in Biothermodynamics, Institute of Biochemistry and Biophysics (IBB), University of Tehran, Tehran, Iran.
authors: Ali A
authors: AA
authors: Moosavi-Movahedi
mesh_terms: D000682:Amyloid; D006801:Humans; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases / Q000737:chemistry* / Q000378:metabolism*; D056004:Molecular Dynamics Simulation; D011487:Protein Conformation; D058767:Protein Unfolding
chemical_list: D000682:Amyloid; C490190:EGLN1 protein, human; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases
keywords: HIF-1; aggregation prone intermediate state; angiogenesis; calorimetry; discrete molecular dynamics; potential of mean force; protein misfolding