Characteristics affecting expression and solubilization of yeast membrane proteins.
Characteristics affecting expression and solubilization of yeast membrane proteins.
复制标题
影响酵母膜蛋白表达和溶解的特性。
DOI：
10.1016/j.jmb.2006.10.004
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发表时间：
2007
影响因子：
5.6
通讯作者：
Dumont,MarkE
中科院分区：
文献类型：
--
作者：
White,MichaelA;Clark,KathleenM;Grayhack,ElizabethJ;Dumont,MarkE
膜蛋白的生物化学和结构分析通常严重依赖于过表达和溶解它们的能力。为了确定真核细胞膜蛋白的特性，可能是成功的过表达的预测，我们分析了超过1000个预测的膜蛋白在最近完成的酿酒酵母蛋白表达文库的基因组互补的表达水平。我们检测到高膜蛋白表达和蛋白质性质，如大小，总体疏水性，跨膜螺旋的数量，和跨膜片段的氨基酸组成之间的统计学显着的正相关和负相关。虽然膜和可溶性蛋白的表达水平表现出类似的整体疏水性负相关，高水平的膜蛋白表达与预测的跨膜段的疏水性呈正相关。为了进一步表征酵母膜蛋白作为结构测定的潜在目标，我们测试了122种表达最高的酵母膜蛋白在六种常用洗涤剂中的溶解度。几乎所有的蛋白质测试可以使用少量的洗涤剂溶解。在某些洗涤剂中的溶解度取决于蛋白质的大小、跨膜片段的数量和预测的跨膜片段的疏水性。这些结果表明，生物信息学的方法可能能够识别膜蛋白，是最适合过表达和洗涤剂溶解的结构和生化分析。生物信息学方法也可以用于重新设计本质上不适合此类研究的蛋白质。
Biochemical and structural analysis of membrane proteins often critically depends on the ability to overexpress and solubilize them. To identify properties of eukaryotic membrane proteins that may be predictive of successful overexpression, we analyzed expression levels of the genomic complement of over 1000 predicted membrane proteins in a recently completed Saccharomyces cerevisiae protein expression library. We detected statistically significant positive and negative correlations between high membrane protein expression and protein properties such as size, overall hydrophobicity, number of transmembrane helices, and amino acid composition of transmembrane segments. Although expression levels of membrane and soluble proteins exhibited similar negative correlations with overall hydrophobicity, high-level membrane protein expression was positively correlated with the hydrophobicity of predicted transmembrane segments. To further characterize yeast membrane proteins as potential targets for structure determination, we tested the solubility of 122 of the highest expressed yeast membrane proteins in six commonly used detergents. Almost all the proteins tested could be solubilized using a small number of detergents. Solubility in some detergents depended on protein size, number of transmembrane segments, and hydrophobicity of predicted transmembrane segments. These results suggest that bioinformatic approaches may be capable of identifying membrane proteins that are most amenable to overexpression and detergent solubilization for structural and biochemical analyses. Bioinformatic approaches could also be used in the redesign of proteins that are not intrinsically well-adapted to such studies.