title: Analyzing folding and degradation of metabolically labelled polypeptides by conventional and diagonal sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
abstract: Efficient protein folding and quality control are essential for unperturbed cell viability. Defects in these processes may lead to production of aberrant polypeptides that are either degraded leading to "loss-of-function" phenotypes, or deposited in or outside cells leading to "gain-of-toxic-function" phenotypes. Elucidation of molecular mechanisms regulating folding and quality control of newly synthesized polypeptides is therefore of greatest interest. Here we describe protocols for metabolic labelling of transfected/infected mammalian cells with [(35)S]-methionine and [(35)S]-cysteine, for immunoisolation from detergent extracts of the selected model proteins and for the investigation of the model polypeptide's intracellular fate in response to chaperone-deletions or to cell exposure to folding or degradation inhibitors.
authors: Institute for Research in Biomedicine, Università della Svizzera italiana-USI, CH-6500, Bellinzona, Switzerland.
authors: Tatiana
authors: T
authors: Soldà
authors: Silvia
authors: S
authors: Olivari
authors: Maurizio
authors: M
authors: Molinari