Expression, purification, and crystallization of neuro- and cytoglobin.
Neuroglobin and cytoglobin, members of the globin family, are present in vertebrate cells at very low concentrations. As the function of both proteins is still a matter of debate, it is very important to be able to produce and purify these proteins, and in general all members of the globin family, to homogeneity. For this purpose, this chapter describes the expression of neuro- and cytoglobin by E. coli and its preparative purification. These proteins are then used in crystallization experiments. Also an analytical purification strategy is discussed in detail.
Dewilde
Sylvia
S
Department of Biomedical Sciences, University of Antwerp, Antwerp, Belgium.
Mees
Kirsten
K
Kiger
Laurent
L
Lechauve
Christophe
C
Marden
Michael C
MC
Pesce
Alessandra
A
Bolognesi
Martino
M
Moens
Luc
L
D000180:Acrylic Resins; D002847:Chromatography, Agarose; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D003460:Crystallization; D000078842:Cytoglobin; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D005914:Globins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D000078787:Neuroglobin; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012685:Sepharose
D000180:Acrylic Resins; C000627265:CYGB protein, human; D000078842:Cytoglobin; D009419:Nerve Tissue Proteins; D000078787:Neuroglobin; D011994:Recombinant Proteins; C023049:sephacryl S 200; D005914:Globins; D012685:Sepharose; C049225:Superose 12