Premium
Towards the quantification of the proteostasis capacity of a cell
Author(s) -
Hatters Danny,
Wood Rebecca,
Vöpel Tobias,
Sharma Abhishek,
Ebbinghaus Simon
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.882.4
Subject(s) - proteostasis , protein folding , microbiology and biotechnology , protein aggregation , protein quality , proteome , barnase , cell , biology , chemistry , biophysics , biochemistry , gene , ribonuclease , rna
How proteins are able to remain properly folded with precise control over their location and activity in the highly crowded cellular environment is one of the miracles of life. Quality control networks govern the life of each protein from synthesis and folding to transport and eventual degradation. However, the capacity of the proteostasis network to recover or overcome protein misfolding or aggregation is not fixed. Instead, it is influenced by environmental stresses, controlled by signalling networks and can be severely compromised by pathogenic proteins such as those seen in neurodegenerative diseases. Here, we present a new biosensor‐based approach to quantitate the proteostasis capacity of cells in terms of both protein folding and aggregation; with the goal of measuring how much energy a cell invests to keep the proteome functional. Our work utilises well‐characterised models of protein folding as “bait” for protein quality control machinery to examine how the cell modulates protein folding and aggregation. The foldedness of various mutants of the bait protein (barnase) can be detected by a fluorescent readout and their aggregation state can be observed. These sensors demonstrate a decline in the proteostasis capacity of cell upon over‐expression of misfolded protein.