title: In Vitro Comparison of the Activity Requirements and Substrate Specificity of Human and Triboleum castaneum PINK1 Orthologues.
abstract: Mutations in the gene encoding the mitochondrial kinase PINK1 cause early-onset familial Parkinson's disease. To understand the biological function of PINK1 and its role in the pathogenesis of Parkinson's disease, it is useful to study its kinase activity towards substrates both in vivo and in vitro. For in vitro kinase assays, a purified Triboleum castaneum PINK1 insect orthologue is often employed, because it displays higher levels of activity when compared to human PINK1. We show, however, that the activity requirements, and more importantly the substrate specificity, differ between both orthologues. While Triboleum castaneum PINK1 readily phosphorylates the PINKtide peptide and Histone H1 in vitro, neither of these non-physiological substrates is phosphorylated by human PINK1. Nonetheless, both Tc and human PINK1 phosphorylate Parkin and Ubiquitin, two physiological substrates of PINK1. Our results show that the substrate selectivity differs among PINK1 orthologues, an important consideration that should be taken into account when extrapolating findings back to human PINK1.
authors.affiliation: Center for the Biology of Disease, Flemish Institute for Biotechnology (VIB), Leuven, Belgium.|Center for Human Genetics, Leuven Institute for Neurodegenerative Disorders and University Hospitals Leuven, University of Leuven, Leuven, Belgium.
authors.forename: Liesbeth
authors.initials: L
authors.lastname: Aerts
authors.affiliation: Center for the Biology of Disease, Flemish Institute for Biotechnology (VIB), Leuven, Belgium.|Center for Human Genetics, Leuven Institute for Neurodegenerative Disorders and University Hospitals Leuven, University of Leuven, Leuven, Belgium.
authors.forename: Katleen
authors.initials: K
authors.lastname: Craessaerts
authors.affiliation: Center for the Biology of Disease, Flemish Institute for Biotechnology (VIB), Leuven, Belgium.|Center for Human Genetics, Leuven Institute for Neurodegenerative Disorders and University Hospitals Leuven, University of Leuven, Leuven, Belgium.|Institute of Neurology, University College London, London, United Kingdom.
authors.forename: Bart
authors.initials: B
authors.lastname: De Strooper
authors.affiliation: Center for the Biology of Disease, Flemish Institute for Biotechnology (VIB), Leuven, Belgium.|Center for Human Genetics, Leuven Institute for Neurodegenerative Disorders and University Hospitals Leuven, University of Leuven, Leuven, Belgium.|Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisbon, Portugal.
authors.forename: Vanessa A
authors.initials: VA
authors.lastname: Morais
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D006657:Histones / Q000378:metabolism; D006801:Humans; D019476:Insect Proteins / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D011494:Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity; D014231:Tribolium / Q000201:enzymology*; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism; D014452:Ubiquitins / Q000378:metabolism; D000099202:PTEN-Induced Putative Kinase
chemical_list: D006657:Histones; D019476:Insect Proteins; D011494:Protein Kinases; D044767:Ubiquitin-Protein Ligases; D014452:Ubiquitins; D000099202:PTEN-Induced Putative Kinase; C111567:parkin protein