title: Substrate profiling of IGF-1R and InsR: identification of a potent pentamer substrate.
abstract: Protein kinases are widely recognized as important therapeutic targets due to their involvement in signal transduction pathways. These pathways are tightly controlled and regulated, notably by the ability of kinases to selectively phosphorylate a defined set of substrates. As part of a study on the substrate requirements of Insulin-like Growth Factor 1 Receptor (IGF-1R) and Insulin Receptor (InsR), we evaluated and applied a universal assay system able to monitor the phosphorylation of unlabelled peptides of any length in real time. In contrast to already reported profiling methodologies, we were able to assess the k(cat)/K(M) ratio of peptides as short as tetramers. Notably, we were able to identify an efficient pentamer substrate that exhibited kinetic properties close to those of a 250-amino acid protein derived from IRS-1, a natural substrate of IGF-1R and InsR.
authors: Novartis Institutes for BioMedical Research, Basel, Switzerland.
authors: Julien
authors: J
authors: Chapelat
authors: Frédéric
authors: F
authors: Berst
authors: Andreas L
authors: AL
authors: Marzinzik
authors: Henrik
authors: H
authors: Moebitz
authors: Peter
authors: P
authors: Drueckes
authors: Doriano
authors: D
authors: Fabbro
authors: Joerg
authors: J
authors: Trappe
authors: Dieter
authors: D
authors: Seebach
mesh_terms: D000595:Amino Acid Sequence; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D015335:Molecular Probes / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000235:genetics; D010766:Phosphorylation; D011485:Protein Binding; D017526:Receptor, IGF Type 1 / Q000737:chemistry*; D011972:Receptor, Insulin / Q000737:chemistry / Q000235:genetics; D013379:Substrate Specificity
chemical_list: D015335:Molecular Probes; D010455:Peptides; D017526:Receptor, IGF Type 1; D011972:Receptor, Insulin