Gel electrophoresis studies of bovine brain white matter proteolipid and myelin proteins

Biochemistry ◽  
1974 ◽  
Vol 13 (13) ◽  
pp. 2704-2712 ◽  
Author(s):  
David S. Chan ◽  
Marjorie B. Lees
Keyword(s):  
White Matter ◽  
Gel Electrophoresis ◽  
Bovine Brain ◽  
Myelin Proteins ◽  
Brain White Matter

De-N-acetyllactotriaosylceramide as a Novel Cationic Glycosphingolipid of Bovine Brain White Matter:  Isolation and Characterization†

Biochemistry ◽  
2005 ◽  
Vol 44 (27) ◽  
pp. 9555-9562 ◽  
Author(s):  
Toshiyuki Hikita ◽  
Keiko Tadano-Aritomi ◽  
Naoko Iida-Tanaka ◽  
Ineo Ishizuka ◽  
Senitiroh Hakomori
Keyword(s):  
White Matter ◽  
Bovine Brain ◽  
Brain White Matter ◽  
Isolation And Characterization

scholarly journals Spectrophotometric assay, solubilization and purification of brain 2′:3′-cyclic nucleotide 3′-phosphodiesterase

1980 ◽  
Vol 191 (1) ◽  
pp. 71-82 ◽  
Author(s):  
Y Nishizawa ◽  
T Kurihara ◽  
Y Takahashi
Keyword(s):  
White Matter ◽  
Cyclic Nucleotide ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Spectrophotometric Assay ◽  
Polyacrylamide Gel ◽  
Hexadecyltrimethylammonium Bromide ◽  
Gel Chromatography ◽  
Brain White Matter

1. A spectrophotometric assay of 2′:3′-cyclic nucleotide 3′-phosphodiesterase (EC 3.1.4.37) based on the use of an acid-base indicator and a buffer having identical pKa values is described. The assay is simple and rapid; it was particularly convenient for monitoring the enzyme activity at various stages of purification. 2. Several proteinases were examined for their ability to solubilize 2′:3′-cyclic nucleotide 3′-phosphodiesterase from delipidated brain white matter. Trypsin (EC 3.4.21.4) and elastase (EC 3.4.21.11) appeared to be more effective than the other proteinases examined. Trypsin, however, caused inactivation; elastase was therefore chosen to solubilize 2′:3′-cyclic nucleotide 3′-phosphodiesterase. When a partially purified preparation of 2′:3′-cyclic nucleotide 3′-phosphodiesterase was treated with elastase, 2′:3′-cyclic nucleotide 3′-phosphodiesterase was solubilized nearly quantitatively. Elastatinal, a specific inhibitor of elastase, specifically inhibited the solubilization with elastase. 3. 2′:3′-cyclic nucleotide 3′-phosphodiesterase was purified from bovine brain white matter by: (i) delipidation; (ii) solubilization with hexadecyltrimethylammonium bromide; (iii) gel chromatography on Sepharose; (iv) ethanol precipitation and resolubilization by digestion with elastase; (v) chromatography on DEAE-Sephadex; (vi) affinity chromatography on 8-(6-aminohexyl)amino-2′-AMP-Sepharose. 4. The purified enzyme migrated as a single protein band on polyacrylamide-gel electrophoresis at pH 4.3 and on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis; the estimated mol.wt. in the latter electrophoresis was 27000-31000. Gel filtration of the purified enzyme through Sephadex G-150 indicated a mol.wt. of 31000. Therefore the purified enzyme is a monomer protein with a mol.wt. of approx. 30000.

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