Development of R7BP inhibitors through cross-linking coupled mass spectrometry and integrated modeling.
Poorni R AdikaramJian-Hua ZhangClaire M KittockMritunjay PandeySergio A HassanNicole G LueGuanghui WangMarjan GucekWilliam F SimondsPublished in: Communications biology (2019)
Protein-protein interaction (PPI) networks are known to be valuable targets for therapeutic intervention; yet the development of PPI modulators as next-generation drugs to target specific vertices, edges, and hubs has been impeded by the lack of structural information of many of the proteins and complexes involved. Building on recent advancements in cross-linking mass spectrometry (XL-MS), we describe an effective approach to obtain relevant structural data on R7BP, a master regulator of itch sensation, and its interfaces with other proteins in its network. This approach integrates XL-MS with a variety of modeling techniques to successfully develop antibody inhibitors of the R7BP and RGS7/Gβ5 duplex interaction. Binding and inhibitory efficiency are studied by surface plasmon resonance spectroscopy and through an R7BP-derived dominant negative construct. This approach may have broader applications as a tool to facilitate the development of PPI modulators in the absence of crystal structures or when structural information is limited.
Keyphrases
- protein protein
- mass spectrometry
- small molecule
- liquid chromatography
- high resolution
- randomized controlled trial
- high performance liquid chromatography
- multiple sclerosis
- capillary electrophoresis
- gas chromatography
- transcription factor
- machine learning
- artificial intelligence
- electronic health record
- deep learning
- binding protein
- dna binding
- tandem mass spectrometry