Co-expression of ferrochelatase allows for complete heme incorporation into recombinant proteins produced in E. coli.
Co-expression of ferrochelatase allows for complete heme incorporation into recombinant proteins produced in E. coli.
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DOI：
10.1016/j.pep.2010.03.010
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发表时间：
2010-09
影响因子：
1.6
通讯作者：
Crane, Brian R.
中科院分区：
文献类型：
--
作者：
Sudhamsu, Jawahar;Kabir, Mariam;Airola, Michael V.;Patel, Bhumit A.;Yeh, Syun-Ru;Rousseau, Denis L.;Crane, Brian R.
Over-expression of heme binding proteins in E. coli often results in sub-optimal heme incorporation and the amount of heme-bound protein produced usually varies with the protein of interest. Complete heme incorporation is important for biochemical characterization, spectroscopy, structural studies, and for the production of homogeneous commercial proteins with high activity. We have determined that recombinant proteins expressed in E. coli often contain less than a full complement of heme because they rather are partially incorporated with free-base porphyrin. Porphyrin-incorporated proteins have similar spectral characteristics as the desired heme-loaded targets, and thus are difficult to detect, even in purified samples. We present a straightforward and inexpensive solution to this problem that involves the co-expression of native ferrochelatase with the protein of interest. The method is shown to be effective for proteins that contain either Cys- or His- ligated hemes.
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影响因子：
1.6
作者：
Varnado, CL;Goodwin, DC
通讯作者：
Goodwin, DC
影响因子：
3.9
作者：
Heinemann, Ilka U.;Jahn, Martina;Jahn, Dieter
通讯作者：
Jahn, Dieter
影响因子：
15
作者：
Davydov, Roman;Sudhamsu, Jawahar;Hoffman, Brian M.
通讯作者：
Hoffman, Brian M.
DOI：
10.1111/j.1432-1033.1991.tb16357.x
发表时间：
1991-11-15
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
ISHIKAWA, K;SATO, M;YOSHIDA, T
通讯作者：
YOSHIDA, T
影响因子：
2.9
作者：
Pant, K;Bilwes, AM;Crane, BR
通讯作者：
Crane, BR