title: Considerations and protocols for the synthesis of custom protein labeling probes.
abstract: Chemists and biologists have long recognized small molecule probes as powerful tools for functional genomics and proteomics studies. The possibility of specifically attaching chemical probes to individual proteins with spatial and temporal resolution has greatly improved our ability to visualize and characterize proteins in their native environment. The continued development of novel molecular probes for protein labeling is, therefore, of fundamental importance to gain new insights into biological processes in living cells and organisms. Several excellent approaches for the site-specific labeling of fusion proteins with chemical probes exist. Herein I discuss the design and generation of chemical probes for the SNAP-tag and CLIP-tag systems. The first part of this chapter is dedicated to reviewing the principles of the SNAP-tag technology, followed by a section dedicated to the development of chemical probes for unique applications, such as super-resolution imaging, protein trafficking and recycling, protein-protein interactions, and biomolecular sensing. The last part of the chapter contains experimental protocols and technical notes for the synthesis of selected SNAP-tag substrates and labeling of SNAP-tag fusion proteins in vitro and in living cells.
authors.affiliation: New England Biolabs, Inc., 240 County Road, Ipswich, MA, 01938, USA, correa@neb.com.
authors.forename: Ivan R
authors.initials: IR
authors.lastname: Corrêa
mesh_terms: D000818:Animals; D015374:Biosensing Techniques; D003432:Cross-Linking Reagents / Q000737:chemistry; D006801:Humans; D015335:Molecular Probes / Q000737:chemistry*; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D013194:Staining and Labeling
chemical_list: D003432:Cross-Linking Reagents; D015335:Molecular Probes; D011993:Recombinant Fusion Proteins