Purification of P0 myelin glycoprotein by a Cu2+-immobilized metal affinity chromatography

Citation
J. Sedzik et al., Purification of P0 myelin glycoprotein by a Cu2+-immobilized metal affinity chromatography, NEUROCHEM R, 24(6), 1999, pp. 723-732
Citations number
65
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Neurosciences & Behavoir
Journal title
NEUROCHEMICAL RESEARCH
ISSN journal
0364-3190 → ACNP
Volume
24
Issue
6
Year of publication
1999
Pages
723 - 732
Database
ISI
SICI code
0364-3190(199906)24:6<723:POPMGB>2.0.ZU;2-K
Abstract
P0 is an abundant myelin glycoprotein of peripheral nerves of vertebrates. Various point mutations of this protein are responsible for hereditary neur opathies. In this paper we described purification of P0 glycoprotein using SDS and a metal chelate affinity chromatography. Purified myelin fraction f rom bovine spinal roots in 0.5% SDS, 0.5 M NaCl, 50 mM Tris-HCl, pH 7.4 is filtered and applied directly to the Cu2+-immobilized affinity chromatograp hy column, equilibrated with the same buffer. After eluting a void volume ( or pass through) fraction, P0 protein was eluted by the same buffer but wit hout salt. To remove contamination from the eluent, further purification is continued on a Concanavalin-A coupled agarose column. We purify within two days, 30 mg of P0 protein of apparent molecular weight 27 kDa. The method can be used to purify recombinant or mutated P0 protein found in severe pat hologies.