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Updated: Aug 10, 2026
Purification of Mouse Brain Vessels
Published on: November 10, 2015
Purification of PASII/PMP22--an extremely hydrophobic glycoprotein of PNS myelin membrane
J Sedzik1, Y Kotake, K Uyemura
1Department of Physiology, Keio University School of Medicine, Tokyo, Japan.
Abstract:
PASII/PMP22 protein (mol. wt 22 kDa, pI 8.8) is an abundant and extremely hydrophobic glycoprotein of PNS myelin which is solubilized effectively by SDS. In humans, this protein is involved in hereditary neuropathies. Here we describe a simple method for purification of PASII/PMP22, suitable for crystallization trials. We usually obtained 10-20 mg PASII/PMP22 from 10 g bovine spinal roots. It is notable, that the original protocol was designated for purification of P0 myelin glycoprotein, and purification of PASII/PMP22 is a bonus. Extensive crystallization trials are in progress.
Insights
We developed a simple method to purify the peripheral nervous system myelin protein 22 (PMP22), crucial for understanding hereditary neuropathies. This technique yields significant amounts of PMP22 for further structural studies.
Area of Science:
- Biochemistry
- Neuroscience
- Structural Biology
Background:
- Peripheral Nervous System (PNS) myelin contains abundant, hydrophobic glycoproteins like PASII/PMP22.
- Mutations in PMP22 are linked to hereditary neuropathies in humans.
- Existing purification methods may not be optimal for PMP22 crystallization.
Purpose of the Study:
- To establish a straightforward and efficient method for purifying PASII/PMP22.
- To obtain sufficient quantities of purified PASII/PMP22 for crystallization trials.
- To facilitate structural and functional studies of PMP22.
Main Methods:
- Utilized a modified protocol originally designed for P0 myelin glycoprotein purification.
- Employed SDS for effective solubilization of the hydrophobic PASII/PMP22.
- Purified PASII/PMP22 from bovine spinal roots.
Main Results:
- Achieved a simple purification protocol yielding 10-20 mg of PASII/PMP22 from 10 g of bovine spinal roots.
- PASII/PMP22 purification was an advantageous outcome of the P0 glycoprotein protocol.
- The purified protein is suitable for subsequent crystallization trials.
Conclusions:
- A practical method for PASII/PMP22 purification has been developed.
- This method provides ample protein for crystallization, advancing structural studies.
- Further research on PMP22 structure may elucidate mechanisms of hereditary neuropathies.
