Purification of the 90 kDa heat shock protein (hsp90) and simultaneous purification of hsp70/hsc70, hsp90 and hsp96 from mammalian tissues and cells using thiophilic interaction chromatography
| Autor: | Yulia Evdokimovskaya, Oleg S. Morenkov, Y. Y. Skarga, V. V. Vrublevskaya |
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| Rok vydání: | 2009 |
| Předmět: |
Affinity matrix
Cells Clinical Biochemistry Biochemistry Chromatography Affinity Analytical Chemistry Sepharose Antigens Neoplasm Heat shock protein Drug Discovery Animals HSP70 Heat-Shock Proteins HSP90 Heat-Shock Proteins Sulfhydryl Compounds Sulfones Animal species Molecular Biology Mercaptoethanol Mammals Pharmacology Chromatography biology Chemistry General Medicine Hsp90 Hsp70 Molecular Weight Organ Specificity Yield (chemistry) biology.protein |
| Zdroj: | Biomedical Chromatography. 23:1208-1216 |
| ISSN: | 1099-0801 0269-3879 |
| DOI: | 10.1002/bmc.1245 |
| Popis: | Heat shock proteins (HSPs) hsp70/hsc70, hsp90 and hsp96 were separated from mammalian cells and tissues on a gel obtained by the reaction of β-mercaptoethanol with divinyl sulfone-activated Sepharose CL-6B (thiophilic gel or T-gel). Hsp90 revealed a much higher affinity towards the T-gel than the other HSPs. One-step thiophilic interaction chromatography of proteins resulted in a more than 80% purity and 85% yield of hsp90. Based on this observation, a simple and efficient method for the purification of hsp90 and a procedure for the simultaneous purification of several HSPs (hsp70/hsc70, hsp90 and hsp96) using thiophilic interaction chromatography was developed. All the HSPs were recovered with a high yield and purity (90–99%). The results indicated that the thiophilic gel is a highly efficient affinity matrix for the purification of hsp90 and can be used in the protocols of purification of different HSPs from cells and tissues of various animal species. Copyright © 2009 John Wiley & Sons, Ltd. |
| Databáze: | OpenAIRE |
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