Incorporation of selenomethionine into induced intracytoplasmic membrane proteins of Rhodobacter species.
Incorporation of selenomethionine into induced intracytoplasmic membrane proteins of Rhodobacter species.
复制标题
将硒代蛋氨酸掺入红杆菌属诱导的胞质内膜蛋白中。
DOI：
10.1007/s10969-005-1936-3
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Hanson,DeborahK
中科院分区：
文献类型：
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作者：
Laible,PhilipD;Hata,AaronN;Crawford,AdamE;Hanson,DeborahK
Efficient multiple- or single-wavelength anomalous dispersion (MAD/SAD) techniques that use tunable X-ray sources at third-generation synchrotrons exploit the anomalous scattering of certain heavy atoms for determination of experimental phases. Development of methods for thein vivosubstitution of methionine by selenomethionine (SeMet) has revolutionized the process for determination of structures of soluble proteins in recent years. Herein, we report methods for biosynthetic incorporation of SeMet into induced intracytoplasmic membrane proteins of two species of theRhodobactergenus of purple non-sulfur photosynthetic bacteria. Amino acid analysis of a membrane protein complex that was purified to homogeneity determined that the extent of SeMet incorporation was extensive and approached quantitative replacement. Diffraction-quality crystals were obtained from SeMet-labeled membrane proteins purified from 2 l of culture. These methods augment the potential utility of photosynthetic bacteria and their inducible membrane systems for the production of foreign membrane proteins for structure determination.
