【摘 要】
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Critical for significant advances in treating a wide range of diseases is the generation of membrane protein samples for use in drug discovery research.Historically,this field has suffered due to diff
【机 构】
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Biosciences Division Argonne National Laboratory USA
【出 处】
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2008中国深圳蛋白质和多肽科学大会
论文部分内容阅读
Critical for significant advances in treating a wide range of diseases is the generation of membrane protein samples for use in drug discovery research.Historically,this field has suffered due to difficulties in obtaining adequate quantities of purified membrane proteins in native form.We describe herein a novel membrane protein expression system,based on the Rhodobacter species of photosynthetic bacteria,which is characterized by an inducible intracytoplasmic membrane.(ICM).This Rhodobacter-based strategy overcomes the major limitation of E.coli-based approaches,which is that native E.coli strains do not induce new membrane concomitantly with protein synthesis to accommodate heterologous membrane proteins and,thus,high-level overexpression in E.coli often results in aggregation and precipitation of incompletely-folded polypeptides.In sharp contrast,these aggregates (or inclusion bodies) have not been observed in any cellular fractions of Rhodobacter expression swains.To further bias the equilibrium towards the production of the folded,functional state of the target proteins,the kinetics of semi-aerobic (or photosynthetic) growth and ICM induction of Rhodobacter can be manipulated to take full advantage of the organisms metabolic diversity to control the timing and extent of membrane protein expression.
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