Purification of the stress protein αB-crystallin and its differentially phosphorylated forms
article
The stress protein α B-crystallin was recently identified as a component of central nervous system myelin that is strongly immunogenic to human T cells. Stress-induced αB-crystallin that accumulates in the central nervous system is phosphorylated and recent evidence indicates that both rodent and human T cells can discriminate between differentially phosphorylated forms of αB-crystallin. For immunological studies, therefore, the availability of purified preparations of αB-crystallin and its various differentially phosphorylated forms would be especially useful. Here we describe a rapid and simple method for the purification of αB-crystallin from adult bovine eye lenses by a combination of size-exclusion chromatography and reversed-phase high-performance liquid chromatography. This yields a preparation of purified α B-crystallin that contains all the various differentially phosphorylated forms of the protein. Subsequent anion- exchange chromatography under denaturing conditions permits the separation of these phosphorylated forms of αB-crystallin into purified fractions with a defined number of phosphorylated serines.
Topics
TNO Identifier
234754
ISSN
00221759
Source
Journal of Immunological Methods, 221(1-2), pp. 159-168.
Pages
159-168
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