The MyoLab uses a multitude of techniques, ranging from spectrophotometric and fluorometric assays by means of kinetic enzyme assays, histochemistry or Western blotting to study metabolism and signalling.
Metabolites
Metabolites, being either a substrate or a product, within tissue or blood, are essential components that can harbour vital information on the physiological processes. Concerning skeletal muscle, our laboratory frequently analyse muscle samples for glucose, glycogen, total creatine and phosphocreatine, and lactate content.
With our collaborators, we are able to use more sophisticated analyses tools (e.g. metabolomics and proteomics) to study other metabolites of interest.
Enzyme activities
Each enzyme of the metabolic pathways, has a specific rate at which it can produce ATP from the fuels mentioned, which describes its activity. However, this rate can be faster or slower from allosteric activation or inhibition, interactions with substrates, activation by other enzymes, or temperature
The following enzyme assays are routinely performed in our lab, each providing information on the flux capacity through a specific pathway:
- 3-hydroxyacetyl Co A dehydrogenase
- creatine kinase
- citrate synthase
- glucose 6 phosphate dehydrogenase
- hexokinase
- lactate dehydrogenase
- phosphofructokinase
- phosphorylase
We are able to develop other enzyme assays of interest.
Metabolite transporters and signalling proteins
Transporters and signalling molecules play important roles in metabolite uptake from the blood. In glucose uptake, the GLUT4 transporters play an important role to lower blood glucose after you have eaten, but also to increase glucose uptake during exercise. We also study the signalling molecules responsible for activating glucose transport, such as the insulin receptor complex, or those during exercise.
We primarily employ Western blotting with antibodies directed against these specific transporters and signalling proteins to study them. With our collaborators, we are able to use more sophisticated analyses tools including GC-MS and nuclear NMR spectrometry.