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JKMRS Volume 23, No 4, pp 104, Validation of protein refolding...
2019년 12월 24일 / 조회수: 541

Validation of protein refolding via 1-dimensional 1H-15N heteronuclear single quantum correlation experiments

 

 

Boram Kim, Joonhyeok Choi, and Kyoung-Seok Ryu*

 

 

Research center for bioconvergence analysis, Korea Basic Science Institute Ochang center,
162 Yeongudanji-Ro, Ochang-Eup, Cheongju-Si, Chungcheongbuk-Do 28119, Republic of Korea

 

  

 

Received Dec 17, 2019; Revised Dec 19, 2019; Accepted Dec 20, 2019

 


Abstract Many proteins are expressed as an insoluble form during the production using Escherichia coli (E. coli) system. Although various methods are applied to increase their amounts of soluble expression, refolding is the only feasible way to obtain a target protein in some cases. Moreover, protein NMR experiments require 13C/15N-labeled proteins that can only be obtained from E. coli systems in terms of cost and technical difficulty. The finding of appropriate refolding conditions for a target protein is a time-consuming process. In particular, it is very difficult to determine whether the refolded protein has a native structure, when a target protein has no enzymatic activity and its refolding yield is very low. Here, we showed that 1-dimensional 1H-15N heteronuclear single quantum correlation (1D 1H-15N HSQC) experiment can be efficiently used to screen an optimal condition for the refolding of a target protein by monitoring both the structure and concentration of the refolded protein.

 

Keywords Blvrb, dialysis using micro-dialyzer, 1-dimensional HSQC experiment, 15N-labeled protein, protein refolding

 

 

* Address correspondence to: Kyoung-Seok Ryu, Research center for bioconvergence analysis, Korea Basic Science Institute, 162 Yeongudanji-Ro, Ochang-Eup, Cheongju-Si, Chungcheongbuk-Do 2811, Republic of Korea, Tel: 82-43-240-5064; Fax: 82-43-240-5059; E-mail: ksryu@kbsi.re.kr

 


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