ReCLIP (reversible cross-link immuno-precipitation): an efficient method for interrogation of labile protein complexes.

ABSTRACT: The difficulty of maintaining intact protein complexes while minimizing non-specific background remains a significant limitation in proteomic studies. Labile interactions, such as the interaction between p120-catenin and the E-cadherin complex, are particularly challenging. Using the cadherin complex as a model-system, we have developed a procedure for efficient recovery of otherwise labile protein-protein interactions. We have named the procedure "ReCLIP" (Reversible Cross-Link Immuno-Precipitation) to reflect the primary elements of the method. Using cell-permeable, thiol-cleavable crosslinkers, normally labile interactions (i.e. p120 and E-cadherin) are stabilized in situ prior to isolation. After immunoprecipitation, crosslinked binding partners are selectively released and all other c

SUBMITTER: Smith AL 

PROVIDER: S-EPMC3024417 | biostudies-literature | 2011 Jan

REPOSITORIES: biostudies-literature