Abstract
We have investigated protein-protein interactions in live Bacillus subtilis cells (a bacterium). B. subtilis’ natural habitat is the soil and the roots of plants, but also the human microbiota. B. subtilis is used worldwide as a model organism.
Unlike eukaryotic cells, bacteria do not have organelles and therefore it was thought that the interior of bacteria is an unorganized mess. But bacteria do have a ‘skelet’: the cytoskeleton. More and more is known about the organization inside bacteria and it turns out that many proteins have a specific location in the cell. Protein complexes might be the bacterial equivalent of eukaryotic organelles.
Earlier research suggests that protein complexes have a transient nature; they are formed when necessary and fall apart afterwards. During this research we studied the dynamics of a few protein complexes; of some proteins involved in sugar metabolism. This has been studied here in live bacteria with a miroscope.
The method that we used to study protein-protein interactions is called FRET (Förster Resonance Energy Transfer). The principle of FRET can be explained with two stem forks. When one stem fork is excited it starts to vibrate; thereby spreading a wave. A second stem fork can absorb this wave if it is nearby and will start to vibrate with the same frequency.
We have labeled sugar metabolism proteins with green and red fluorescent proteins (‘our two stem forks’). If we can observe the red protein after exciting the green protein, then we have proven that the sugar metabolism proteins interact.
Unlike eukaryotic cells, bacteria do not have organelles and therefore it was thought that the interior of bacteria is an unorganized mess. But bacteria do have a ‘skelet’: the cytoskeleton. More and more is known about the organization inside bacteria and it turns out that many proteins have a specific location in the cell. Protein complexes might be the bacterial equivalent of eukaryotic organelles.
Earlier research suggests that protein complexes have a transient nature; they are formed when necessary and fall apart afterwards. During this research we studied the dynamics of a few protein complexes; of some proteins involved in sugar metabolism. This has been studied here in live bacteria with a miroscope.
The method that we used to study protein-protein interactions is called FRET (Förster Resonance Energy Transfer). The principle of FRET can be explained with two stem forks. When one stem fork is excited it starts to vibrate; thereby spreading a wave. A second stem fork can absorb this wave if it is nearby and will start to vibrate with the same frequency.
We have labeled sugar metabolism proteins with green and red fluorescent proteins (‘our two stem forks’). If we can observe the red protein after exciting the green protein, then we have proven that the sugar metabolism proteins interact.
| Original language | English |
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| Qualification | Doctor of Philosophy |
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| Supervisors/Advisors |
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| Award date | 11-Dec-2015 |
| Place of Publication | [Groningen] |
| Publisher | |
| Print ISBNs | 978-90-367-8308-8 |
| Electronic ISBNs | 978-90-367-8307-1 |
| Publication status | Published - 2015 |