In-cell destabilization of a homodimeric protein complex detected by DEER spectroscopy
Yin Yang, Shen-Na Chen, Feng Yang, Xia-Yan Li, Akiva Feintuch, Xun‐Cheng Su, Daniella Goldfarb
Abstract
was markedly higher in cells than in solution and dilute cell lysate. As expected, this increase was partially recapitulated under conditions of high salt concentrations, given that conserved salt bridges at the dimer interface are critically required for association. Unexpectedly, however, also the addition of the crowding agent Ficoll destabilized the dimer while the addition of bovine serum albumin (BSA) and lysozyme, often used to represent interaction with charged macromolecules, had no effect. Our results highlight the potential of DEER for in-cell study of proteins as well as the complexities of the effects of the cellular milieu on protein structures and stability.