title: Identification of peptidic substrates for the human kinase Myt1 using peptide microarrays.
abstract: Myt1 kinase is a member of the Wee-kinase family involved in G2/M checkpoint regulation of the cell cycle. So far, no peptide substrate suitable for activity-based screening has been reported, hampering systematic development of Myt1 kinase inhibitors. Myt1 inhibitors had to be identified by using either binding assays or activity assays with expensive proteinous substrates. Here, a peptide microarray approach was used to identify peptidic Myt1 substrates. Wee1 kinase was profiled for comparison using the same technology. Myt1 hits from peptide microarray experiments were verified in solution by a fluorescence polarization assay and several peptide substrates derived from cellular proteins were identified. Subsequently, phosphorylation site determination was carried out by MS fragmentation studies and identified substrates were validated by kinase inhibitor profiling.
authors.affiliation: Institute of Pharmacy, Department of Medicinal Chemistry, Martin-Luther-University Halle-Wittenberg, W.-Langenbeck-Str. 4, 06120 Halle, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Rohe
authors.affiliation: Institute of Pharmacy, Department of Medicinal Chemistry, Martin-Luther-University Halle-Wittenberg, W.-Langenbeck-Str. 4, 06120 Halle, Germany.
authors.forename: Charlott
authors.initials: C
authors.lastname: Platzer
authors.affiliation: Institute of Biochemistry and Biotechnology, Department of Enzymology, Martin-Luther-University Halle-Wittenberg, Kurt-Mothes-Str. 3, 06120 Halle, Germany.
authors.forename: Antonia
authors.initials: A
authors.lastname: Masch
authors.affiliation: Institute of Pharmacy, Department of Medicinal Chemistry, Martin-Luther-University Halle-Wittenberg, W.-Langenbeck-Str. 4, 06120 Halle, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Greiner
authors.affiliation: Institute of Pharmacy, Department of Medicinal Chemistry, Martin-Luther-University Halle-Wittenberg, W.-Langenbeck-Str. 4, 06120 Halle, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Henze
authors.affiliation: Institute of Pharmacy, Department of Bioanalytics, Martin-Luther-University Halle-Wittenberg, W.-Langenbeck-Str. 4, 06120 Halle, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Ihling
authors.affiliation: Institute of Pharmacy, Department of Pharmacology, Martin-Luther-University Halle-Wittenberg, W.-Langenbeck-Str. 4, 06120 Halle, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Erdmann
authors.affiliation: Institute of Biochemistry and Biotechnology, Department of Enzymology, Martin-Luther-University Halle-Wittenberg, Kurt-Mothes-Str. 3, 06120 Halle, Germany.
authors.forename: Mike
authors.initials: M
authors.lastname: Schutkowski
authors.affiliation: Institute of Pharmacy, Department of Medicinal Chemistry, Martin-Luther-University Halle-Wittenberg, W.-Langenbeck-Str. 4, 06120 Halle, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Sippl
authors.affiliation: Institute of Pharmacy, Department of Medicinal Chemistry, Martin-Luther-University Halle-Wittenberg, W.-Langenbeck-Str. 4, 06120 Halle, Germany. Electronic address: matthias.schmidt@pharmazie.uni-halle.de.
authors.forename: Matthias
authors.initials: M
authors.lastname: Schmidt
mesh_terms: D000595:Amino Acid Sequence; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D040081:Protein Array Analysis; D013379:Substrate Specificity; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; C078341:MYT1 protein, human; D010455:Peptides; D014157:Transcription Factors
keywords: Fluorescence polarization assay; Kinase substrate profiling; Myt1 kinase; Peptide microarray