Purification process for heat shock proteins using aqueous two-phase system and PEG fractional precipitation

被引：7

作者：

Kuboi, R

Hasegawa, T

Yano, K

Komasawa, I

机构：

[1] Department of Chemical Engineering, Osaka University, Toyonaka

来源：

JOURNAL OF CHEMICAL ENGINEERING OF JAPAN | 1995年 / 28卷 / 06期

关键词：

heat shock protein; aqueous two-phase system; PEG fractional precipitation; surface property; biospecific Affinity;

D O I：

10.1252/jcej.28.797

中图分类号：

TQ [化学工业];

学科分类号：

0817 ;

摘要：

A simple and effective purification process for heat shock proteins (HSPs), in which PEG fractional precipitation was combined with an aqueous two-phase system (ATPS), was successfully developed based upon the proteins' molecular surface properties. Both GroEL and GroES, typical HSPs from E.coli, were selectively partitioned to the PEG (top) phase of the ATPS, GroEL and GroES were selectively and stepwisely concentrated from the above PEG phase by PEG fractional precipitation, In addition, the purification of GroEL and GroES as a complex by using biospecific affinity with adenosine triphosphate (ATP) was achieved. GroEL and GroES can be purified either as individual native molecules or as a 1:1 complexed state by the control of ATP addition.

引用

页码：797 / 802

页数：6

共 17 条

[1]

Albertsson P.A., Partition of Cell Particles and Macromolecules, pp. 112-146, (1986)

[2]

Atha D.H., Ingham K.C., Mechanism of Precipitation of Proteins by Polyethylene Glycols, J. Biol. Chem., 256, pp. 12108-12117, (1981)

[3]

Bradford M.M., A Rapid and Sensitive Method for the Quantification of Micrograin Quantities of Protein Utilizing the Principle of Protein-Dye Binding, Anal. Biochem., 72, pp. 248-254, (1976)

[4]

Chandrasekhar G.N., Tilly K., Woolford C., Hendrix R., Georgopoulos C., Purification and Properties of the groES Morphogenetic Protein of Escherichia coli, J. Biol. Chem., 261, pp. 12414-12419, (1986)

[5]

Fisher M.T., Promotion of the in Vitro Renaturation of Dodecameric Glutamine Synthetase from Escherichia coli in the Presence of GroEL (Chaperonin-60) and ATP, Biochemistry, 31, pp. 3955-3963, (1991)

[6]

Harrison R.G., Protein Purification Process Engineering, pp. 115-208, (1994)

[7]

Hemmingsen S.M., Woolfold C., van der Vies S.M., Tilly K., Dennis D.T., Georgopoulos C.P., Hendrix R.W., Ellis R.J., Homologous Plant and Bacterial Proteins Chaperone Oligomeric Protein Assembly, Nature, 333, pp. 330-334, (1988)

[8]

Hendrix R.W., Purification and Properties of groE, a Host Protein Involved in Bacteriophage Assembly, JMol. Biol., 129, pp. 375-392, (1979)

[9]

Horio T., Theory and Practice on Enzymes and Other Proteins, pp. 93-117, (1994)

[10]

Ingham K.C., Precipitation of Proteins with Polyethylene Glycol, Methods in Enzymology, 182, pp. 301-306, (1990)

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