Characterization of a recombinant murine 18.5-kDa myelin basic protein

被引：69

作者：

Bates, IR

Matharu, P

Ishiyama, N

Rochon, D

Wood, DD

Polverini, E

Moscarello, MA

Viner, NJ

Harauz, G ^{[1
]}

机构：

[1] Univ Guelph, Dept Mol Biol & Genet, Guelph, ON N1G 2W1, Canada

[2] Univ Guelph, Biophys Interdept Grp, Guelph, ON N1G 2W1, Canada

[3] Univ Bristol, Sch Med Sci, Dept Pathol & Microbiol, Bristol BS8 1TD, Avon, England

[4] Hosp Sick Children, Dept Biochem & Struct Biol, Toronto, ON M5G 1X8, Canada

[5] Univ Parma, Ist Nazl Fis Mat, I-43100 Parma, Italy

[6] Univ Parma, Dipartimento Fis, I-43100 Parma, Italy

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2000年 / 20卷 / 02期

基金：

英国惠康基金; 英国医学研究理事会; 加拿大自然科学与工程研究理事会;

关键词：

myelin basic protein; multiple sclerosis; His-tag; mass spectrometry; circular dichroism; electron microscopy; ganglioside; phosphatidylinositol;

D O I：

10.1006/prep.2000.1307

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A recombinant hexahistidine-tagged 18.5-kDa isoform of murine myelin basic protein has been characterized biochemically and immunogenically, by mass spectrometry, by circular dichroism under various conditions (in aqueous solution, with monosialoganglioside G(M1), and in 89% 2-propanol), and by transmission electron microscopy. The preparations of this protein indicated a high degree of purity and homogeneity, with no significant posttranslational modifications, Circular dichroic spectra showed that this preparation had the same degree of secondary structure as the natural bovine 18.5-kDa isoform of myelin basic protein. Incubation of the recombinant protein with lipid monolayers containing a nickel-chelating lipid resulted in the formation of fibrous assemblies that formed paracrystals of spacings 4.8 nm between fibers and 3-4 nm along them. (C) 2000 Academic Press.

引用

页码：285 / 299

页数：15

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