Myelin basic protein purified on an ion-exchange continuous polymer bed in the presence of ethylene glycol and salt possesses activity against p-nitrophenyl acetate.
In this paper we describe a fast and mild method based on the use of a unique cation exchanger and buffers containing ethylene glycol and salt for the purification of the myelin basic protein (MBP; MW 18.5 kDa). MBP thus purified hydrolyses catalytically p-nitrophenyl acetate. This esterase activity facilitates not only the purification of MBP but also indicates that probably it is in its native state, i.e. there is a good chance that the purified molecules are structurally and chemically identical. This is a prerequisite to obtain crystals appropriate for x-ray diffraction and other studies.
Sedzik
J
J
Department of Biochemistry, Uppsala University, Sweden.
Mohammad
J
J
Hjertén
S
S
D009597:4-Nitrophenylphosphatase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002852:Chromatography, Ion Exchange / Q000379:methods; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D004950:Esterases / Q000737:chemistry; D019855:Ethylene Glycol; D005026:Ethylene Glycols; D005260:Female; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D004676:Myelin Basic Protein / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D009596:Nitrophenols; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D012965:Sodium Chloride; D013116:Spinal Cord / Q000378:metabolism*
D003902:Detergents; D005026:Ethylene Glycols; D004676:Myelin Basic Protein; D009596:Nitrophenols; D012965:Sodium Chloride; C008642:4-nitrophenyl acetate; D004950:Esterases; D009597:4-Nitrophenylphosphatase; D019855:Ethylene Glycol