title: Protein fishing with chiral molecular baits.
abstract: Chirality plays a central role in various biological recognition processes. Here a methodology was developed to utilize chiral recognition processes for the selective biotinylation of proteins in crude cell lysates. Two pairs of diastereomeric probes containing benzophenone and biotin were prepared through solid-phase synthesis. Protein-binding selectivity of each probe was examined by photo-cross-linking of cell lysates, followed by SDS-PAGE and Western blot. The study revealed that our approach permits selective labeling of benzophenone-binding proteins in complex proteomes. In addition, it was found that the selectivity depends largely on a single chiral center and substitutions in the vicinity of benzophenone. Taken together, the current work demonstrates that chiral recognition process can be employed to selectively label proteins in complex proteomes. Thus the study opens up the possibility to expand the scope of chemical proteomics research for various applications, including biomarker discovery, drug screening and development.
authors.affiliation: Department of Chemistry, CUNY-Hunter College, New York, New York 10021, USA. akawamur@hunter.cuny.edu
authors.forename: Akira
authors.initials: A
authors.lastname: Kawamura
authors.forename: Sagit
authors.initials: S
authors.lastname: Hindi
mesh_terms: D001710:Biotin / Q000737:chemistry; D045744:Cell Line, Tumor; D003432:Cross-Linking Reagents; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D015336:Molecular Probe Techniques; D015335:Molecular Probes / Q000138:chemical synthesis / Q000737:chemistry*; D009363:Neoplasm Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*; D040901:Proteomics; D021241:Spectrometry, Mass, Electrospray Ionization; D013237:Stereoisomerism
chemical_list: D003432:Cross-Linking Reagents; D015335:Molecular Probes; D009363:Neoplasm Proteins; D011506:Proteins; D001710:Biotin