Equilibrium thrown off balance
Scientists at the Leibniz-Institut für Molekulare Pharmakologie in Berlin have now shown, for the first time, that the ER loses its oxidative power in advanced age, which shifts the reducing/oxidising equilibrium – redox for short – in this compartment. This leads to a decline in the capacity to form the disulphide bridges that are so important for correct protein folding. As a consequence, many proteins can no longer mature properly and become unstable.
Although, it was already known that increased protein misfolding occurs with the progression of ageing, it was not known whether the redox equilibrium is affected. Likewise, it was not known that the loss of oxidative power in the ER also affects the equilibrium in another compartment of the cell: in reverse, namely, the otherwise protein-reducing cytosol becomes more oxidising during ageing, which leads to the known oxidative protein damage such those caused by the release of free radicals.
"Up to now, it has been completely unclear what happens in the endoplasmic reticulum during the ageing process. We have now succeeded in answering this question," says Dr. Janine Kirstein, first author of the study, which has been published in EMBO Journal*. At the same time, the scientists were able to show that there is a strong correlation between protein homoeostasis and redox equilibrium. "This is absolutely new and helps us to understand why secretory proteins become unstable and lose their function in advanced age and after stress. This may explain why the immune response declines as we get older," the biologist explains further.
Stress has the same effects as ageing
The researchers also demonstrated the decline of the oxidative milieu of the ER after stress. When they synthesised amyloid protein fibrils in the cell, which cause diseases such as Alzheimer's, Parkinson's or Huntington's disease, they set the same cascade in motion. Apart from this, they were able to show that amyloids that are synthesised in a certain tissue also have negative effects on the redox equilibrium in another tissue within the same organism. "Protein stress leads to the same effects as ageing," explains Kirstein. "Our findings are thus not only interesting with regards to ageing, but also concerning neurodegenerative diseases such as Alzheimer's."
For their experiments, the team of researchers used nematodes - an established model system for investigating ageing processes on a molecular level. Since the nematode is transparent, the researchers were able to use fluorescence-based sensors in order to measure oxidation in the individual cell compartments. It was thus possible to track precisely in the living nematode how the redox condition changes with advancing age. In addition, the influence of protein aggregation on the redox homeostasis was investigated in cultivated cells of human origin. The data were fully consistent with those from the nematode.
Using the findings to identify new diagnostic biomarkers
"We gained a lot of insight, but have also learned that ageing is much more complex than previously assumed," stresses the biologist Kirstein. Thus, for example, the mechanism of the signal transduction of protein folding stress to the redox equilibrium – both within the cell from one compartment to another and also between two different tissues – remains completely unclear.
Nevertheless, research of ageing has taken a major step forward as a result of the findings from Berlin, particularly since it promises a practical benefit. The redox equilibrium may serve as a basis for new biomarkers for diagnosing both ageing and neurodegenerative processes in the future. Janine Kirstein: "The approach is less likely to be useful for therapeutic purposes at present, but the development of diagnostic tools is certainly conceivable."
The project is a cooperation between laboratories from Berlin, Chicago, Kyoto and Munich.
*Kirstein J, Morito D, Kakihana T, Sugihara M, Minnen A, Hipp MS, Nussbaum-Krammer C, Hartl FU, Nagata K & Morimoto RI. Proteotoxic stress and ageing triggers the loss of redox homeostasis across cellular compartments. EMBO Journal, 2015 in press
The Leibniz-Institut für Molekulare Pharmakologie (FMP) is part of the Forschungsverbund Berlin e.V. (FVB), who legally represents eight non-university research institutes - members of the Leibniz Association - in Berlin. The institutions pursue common interests within the framework of a single legal entity while maintaining their scientific autonomy. More than 1,500 employees work within the research association. The eight institutes were founded in 1992 and emerged from former institutes of the GDR Academy of Sciences.
Text: Beatrice Hamberger, Translation Mick Locke
Dr. Janine Kirstein
Leibniz-Institut für Molekulare Pharmakologie (FMP)
Tel.: 030 94793 250
Tel.: 030 94793 104