Light-Controlled Affinity Purification of Protein Complexes Exemplified by the Resting ZAP70 Interactome.

ABSTRACT: Multiprotein complexes control the behavior of cells, such as of lymphocytes of the immune system. Methods to affinity purify protein complexes and to determine their interactome by mass spectrometry are thus widely used. One drawback of these methods is the presence of false positives. In fact, the elution of the protein of interest (POI) is achieved by changing the biochemical properties of the buffer, so that unspecifically bound proteins (the false positives) may also elute. Here, we developed an optogenetics-derived and light-controlled affinity purification method based on the light-regulated reversible protein interaction between phytochrome B (PhyB) and its phytochrome interacting factor 6 (PIF6). We engineered a truncated variant of PIF6 comprising only 22 amino acids that can be

SUBMITTER: Horner M 

PROVIDER: S-EPMC6399385 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature