JKMRS Volume 23, No 4, pp 104, Validation of protein refolding... | |
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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, 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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첨부파일 | 4_RyuKS.pdf |