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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2011.1
- 수록면
- 279 - 284 (6page)
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초록· 키워드
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The glutathione S-transferase (GST) system is useful for increasing protein solubility and purifying soluble GST fusion proteins.
However, purifying half of the GST fusion proteins is still difficult, because they are virtually insoluble under non-denaturing conditions. To optimize a simple and rapid purification condition for GST-pyruvate kinase muscle 2 (GST-PKM2) protein,we used 1% sarkosyl for lysis and a 1：200 ratio of sarkosyl to Triton X-100 (S-T) for purification. We purified the GST-PKM2 protein with a high yield, approximately 5 mg/L culture, which was 33 times higher than that prepared using a conventional method. Notably, the GST-high-temperature requirement A2 (GST-HtrA2) protein, used as a model protein for functional activity, fully maintained its proteolytic activity,even when purified under our S-T condition. This method may be useful to apply to other biologically important proteins that become highly insoluble in the prokaryotic expression system.
[BMB reports 2011; 44(4): 279-284]
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참고문헌 (23)
참고문헌 신청
Frangioni, J. V / 1993 / Solubilization and purification of enzymatically active glutathione S-transferase (pGEX) fusion proteins / Anal. Biochem 210 : 179 ~ 187
Frankel, S / 1991 / The use of sarkosyl in generating soluble protein after bacterial expression / Proc. Natl. Acad. Sci 88 : 1192 ~ 1196
Hahn, A. W / 1993 / Expression of soluble and insoluble fibronectin in rat aorta: effects of angiotensin II and endothelin- 1 / Biochem. Biophys. Res. Commun 192 : 189 ~ 197
Harper, S / 2011 / Purification of proteins fused to glutathione S-transferase / Methods Mol. Biol 681 : 259 ~ 280
Hartman, J / 1992 / Affinity purification of insoluble recombinant fusion proteins containing glutathione- S-transferase / Biotechnol. Bioeng 39 : 828 ~ 832
Frankel, S / 1991 / The use of sarkosyl in generating soluble protein after bacterial expression / Proc. Natl. Acad. Sci 88 : 1192 ~ 1196
Hahn, A. W / 1993 / Expression of soluble and insoluble fibronectin in rat aorta: effects of angiotensin II and endothelin- 1 / Biochem. Biophys. Res. Commun 192 : 189 ~ 197
Harper, S / 2011 / Purification of proteins fused to glutathione S-transferase / Methods Mol. Biol 681 : 259 ~ 280
Hartman, J / 1992 / Affinity purification of insoluble recombinant fusion proteins containing glutathione- S-transferase / Biotechnol. Bioeng 39 : 828 ~ 832
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