Proteomic mapping provides powerful insights into functional myelin biology - PubMed (original) (raw)

Proteomic mapping provides powerful insights into functional myelin biology

Christopher M Taylor et al. Proc Natl Acad Sci U S A. 2004.

Abstract

Myelin is a dynamic, functionally active membrane necessary for rapid action potential conduction, axon survival, and cytoarchitecture. The number of debilitating neurological disorders that occur when myelin is disrupted emphasizes its importance. Using high-resolution 2D gel electrophoresis, mass spectrometry, and immunoblotting, we have developed an extensive proteomic map of proteins present in myelin, identifying 98 proteins corresponding to at least 130 of the approximately 200 spots on the map. This proteomic map has been applied to analyses of the localization and function of selected proteins, providing a powerful tool to investigate the diverse functions of myelin.

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Figures

Fig. 1.

Fig. 1.

2D PAGE mapping of myelin proteins. Myelin proteins were separated by using a pH 3-10 IPG strip and 12% SDS/PAGE gels, and the gels stained with silver nitrate. Proteins have been assigned numbers corresponding to the spot numbers listed in Table 1. Approximate molecular mass (kDa) and pI values are listed.

Fig. 2.

Fig. 2.

2D immunoblotting of myelin proteins. Each numbered immunoblot is shown above its corresponding region from a silver-stained gel. Numbers correspond to protein spots from Fig. 1 and Table 1. For display purposes, OSP/claudin-11 and proteolipid protein/DM20 immunoblots and silver-stained gel regions were minimized relative to other spots.

Fig. 3.

Fig. 3.

NSF is a previously undescribed component of Schwann cell microvilli. (a) 2D PAGE of adult PNS myelin proteins shows the presence of NSF (arrows). (Inset) 1D immunoblot of NSF in brain homogenate (B) and sciatic nerve (SN). (b and c) Immunostaining of NSF (Left and Right in green) and voltage-gated sodium channels (Center and Right in red) in adult sciatic nerve transverse section (b) and cross section (c). (Bar = 3 μm.) (d) Immunostaining of NSF (Left and Right in green) and moesin (Center and Right in red) in adult sciatic nerve cross section. (Bar = 3 μm.) (e) Immunostaining of postnatal day 1 sciatic nerve transverse section. (Bar = 2 μm.) Axon orientation in b is from bottom left to upper right. Axon orientation in e is from left to right.

Fig. 4.

Fig. 4.

Nm23A and -B are dramatically reduced in CGT-null myelin. (a and b) Myelin proteins purified from wild-type (a) and CGT-null mice (b) were separated by 2D PAGE and silver stained. Nm23A (2) and -B (3) levels were dramatically reduced in CGT-null myelin when compared with wild-type myelin. (c) Similar reductions also seen in brain homogenates (BH) were confirmed by immunoblotting. Note the enrichment of Nm23 in myelin (My) compared with brain homogenate. (d) Levels of actin remained unchanged in CGT-null mice compared with wild-type controls.

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References

    1. Pfeiffer, S. E., Warrington, A. E. & Bansal, R. (1993) Trends Cell Biol. 3, 191-197. - PubMed
    1. Keegan, B. M. & Noseworthy, J. H. (2002) Annu. Rev. Med. 53, 285-302. - PubMed
    1. Koeppen, A. H. & Robitaille, Y. (2002) J. Neuropathol. Exp. Neurol. 61, 747-759. - PubMed
    1. Zhou, L. & Griffin, J. W. (2003) Curr. Opin. Neurol. 16, 307-313. - PubMed
    1. Morell, P. & Norton, W. T. (1980) Sci. Am. 242 (5), 88-90, 92, 96. - PubMed

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