双光子荧光相关光谱法和分子亮度分析显示酵母核糖体柄在体内的异质性
Yeast ribosomal stalk heterogeneity in vivo shown by two-photon FCS and molecular brightness analysis.
作者信息
García-Marcos Alberto, Sánchez Susana A, Parada Pilar, Eid John, Jameson David M, Remacha Miguel, Gratton Enrico, Ballesta Juan P G
机构信息
Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid, 28049 Madrid, Spain.
出版信息
Biophys J. 2008 Apr 1;94(7):2884-90. doi: 10.1529/biophysj.107.121822. Epub 2007 Dec 20.
Abstract
The stalk of Saccharomyces cerevisiae ribosomes contains, on average, five distinct proteins, namely P0 and four acidic proteins, P1alpha, P1beta, P2alpha, and P2beta. Each ribosome contains only one copy of P0, but the distribution of the acidic proteins among the ribosome population in vivo has not been determined. Using two-photon fluorescence correlation spectroscopy and scanning FCS, on cells expressing EGFP-tagged P0, P1, and P2 proteins, we show, with brightness analysis, that individual yeast ribosomes in vivo are compositionally heterogeneous in regard to P1alpha, P1beta, P2alpha, and P2beta. These results are relevant to the hypothesis, based on in vitro studies, that the overall cellular pattern of expressed proteins can be determined by the distribution of the stalk proteins among the ribosome population.
摘要
酿酒酵母核糖体的柄部平均含有五种不同的蛋白质，即P0和四种酸性蛋白质P1α、P1β、P2α和P2β。每个核糖体仅含有一份P0，但体内核糖体群体中酸性蛋白质的分布尚未确定。利用双光子荧光相关光谱法和扫描荧光相关光谱法，对表达EGFP标记的P0、P1和P2蛋白的细胞进行分析，通过亮度分析我们发现，体内单个酵母核糖体在P1α、P1β、P2α和P2β方面在组成上是异质的。这些结果与基于体外研究的假设相关，即表达蛋白的整体细胞模式可由柄部蛋白在核糖体群体中的分布决定。
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