title: A comparative study of ATP analogs for phosphorylation-dependent kinase-substrate crosslinking.
abstract: Kinase-catalyzed protein phosphorylation is an important post-translational modification that regulates a variety of cellular functions. Identification of the many substrates of a specific kinase is critical to fully characterize cell biology. Unfortunately, kinase-substrate interactions are often transient, which makes their identification challenging. Here, the transient kinase-substrate complex was stabilized by covalent crosslinking using γ-phosphate modified ATP analogs. Building upon prior use of an ATP-aryl azide photocrosslinking analog, we report here the creation of an ATP-benzophenone photocrosslinking analog. ATP-benzophenone displayed a higher conversion percentage but more diffuse crosslinking compared to the ATP-aryl azide analog. A docking study was also performed to rationalize the conversion and crosslinking data. In total, the photocrosslinking ATP analogs produced stable kinase-substrate complexes that are suitable for future applications characterizing cell signaling pathways.
authors: Department of Chemistry, Wayne State University, Detroit, MI 48202, United States.
authors: Satish
authors: S
authors: Garre
authors: Department of Chemistry, Wayne State University, Detroit, MI 48202, United States.
authors: Chamara
authors: C
authors: Senevirathne
authors: Department of Chemistry, Wayne State University, Detroit, MI 48202, United States. Electronic address: pflum@wayne.edu.
authors: Mary Kay H
authors: MK
authors: Pflum
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D006801:Humans; D010766:Phosphorylation / Q000502:physiology*; D011494:Protein Kinases / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D015398:Signal Transduction; D013379:Substrate Specificity
chemical_list: D000255:Adenosine Triphosphate; D011494:Protein Kinases
keywords: ATP analogs; Kinase; Kinase substrates; Photocrosslinking