The Yusuf Hamied Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK
Bidesh Mahata
Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge CB10 1SA, UK; EMBL-European Bioinformatics Institute, Wellcome Genome Campus, Hinxton, Cambridge CB10 1SD, UK; Division of Immunology, Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK
Jhuma Pramanik
Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge CB10 1SA, UK; Division of Immunology, Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK
Marco L. Hennrich
Structural and Computational Biology Unit, European Molecular Biology Laboratory, EMBL, Heidelberg, Germany
Anne-Claude Gavin
Structural and Computational Biology Unit, European Molecular Biology Laboratory, EMBL, Heidelberg, Germany; Molecular Medicine Partnership Unit, European Molecular Biology Laboratory, EMBL, Heidelberg, Germany; University of Geneva, Department for Cell Physiology and Metabolism, Centre Medical Universitaire, Rue Michel-Servet 1, CH-1211 Geneva 4, Switzerland
Steven V. Ley
The Yusuf Hamied Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK
Sarah A. Teichmann
Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge CB10 1SA, UK; Theory of Condensed Matter, Cavendish Laboratory, 19 JJ Thomson Avenue, Cambridge CB3 0HE, UK; Corresponding author
Summary: Pregnenolone (P5) promotes prostate cancer cell growth, and de novo synthesis of intratumoural P5 is a potential cause of development of castration resistance. Immune cells can also synthesize P5 de novo. Despite its biological importance, little is known about P5's mode of actions, which appears to be context dependent and pleiotropic. A comprehensive proteome-wide spectrum of P5-binding proteins that are involved in its trafficking and functionality remains unknown. Here, we describe an approach that integrates chemical biology for probe synthesis with chemoproteomics to map P5-protein interactions in live prostate cancer cells and murine CD8+ T cells. We subsequently identified P5-binding proteins potentially involved in P5-trafficking and in P5's non-genomic action that may drive the promotion of castrate-resistance prostate cancer and regulate CD8+ T cell function. We envisage that this methodology could be employed for other steroids to map their interactomes directly in a broad range of living cells, tissues, and organisms.
