MBP2 (Myelin Basic Protein Squared) is a science outreach project I was involved in during my postdoc at the University of Kent in a local school that has now completed. The outreach project studied the formation of the myelin sheath caused by phosphorylation of MBP. This has been a successful project engaging with students and involving a wide variety of biology from genetic transformation & protein expression, protein extraction, protein & DNA analysis and biophysics. The project has received Wellcome Trust funding in the past with maintained support from the University of Kent School of Biosciences and more recently the School of Physical Sciences.
Myelin basic protein is importing in forming the insulting layer around neurons, the myelin sheath. Without the myelin sheath, neurons can trigger action potential randomly, a condition called Multiple sclerosis, which can lead to serious disabilities1.
The project focused on measuring whether the purified MBP protein variants embedded within the lipid membrane. If the protein did not embed into the lipid monolayer, it could suggest the protein is responsible for the myelin sheath not forming around neurons. The monolayer was made by forming lipid micelles and setting them onto a Quartz Crystal Microbalance (QCM). The QCM can measure nanogram changes by the dampening the quartz frequency defined as the Sauerbrey equation2. Each stage of the MBP application can be measured including the final wash stage where if the weight decreased, it implies the loss of MBP and lack of embedding within the lipid monolayer.
Furthermore, we can use the mass change to calculate the molecule per area (molecule/cm2) or area per molecule (cm2/molecule). It is important we have a lipid monolayer formed as well as calculating how many myelin basic protein derivatives have affinity for the layer. We know the weight of a molecule based on the kilodalton (kDa) to calculate the density.