title: 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.
abstract: Heat shock proteins (HSPs) hsp70/hsc70, hsp90 and hsp96 were separated from mammalian cells and tissues on a gel obtained by the reaction of beta-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.
authors.affiliation: Laboratory of Cell Cultures and Cellular Engineering, Institute of Cell Biophysics, Russian Academy of Sciences, Pushchino 142290, Russia. yyskarga@gmail.ru
authors.forename: Yuri
authors.initials: Y
authors.lastname: Skarga
authors.forename: Veronika
authors.initials: V
authors.lastname: Vrublevskaya
authors.forename: Yulia
authors.initials: Y
authors.lastname: Evdokimovskaya
authors.forename: Oleg
authors.initials: O
authors.lastname: Morenkov
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002477:Cells; D002846:Chromatography, Affinity / Q000379:methods*; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D018841:HSP90 Heat-Shock Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008322:Mammals / Q000378:metabolism*; D008623:Mercaptoethanol / Q000737:chemistry; D008970:Molecular Weight; D009928:Organ Specificity; D012685:Sepharose / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D013450:Sulfones / Q000737:chemistry
chemical_list: D000951:Antigens, Neoplasm; D018840:HSP70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; D013438:Sulfhydryl Compounds; D013450:Sulfones; C052646:sarcoma glycoprotein gp96 rejection antigens; C009873:divinyl sulfone; D008623:Mercaptoethanol; D012685:Sepharose