Bottlenecks and roadblocks in high-throughput XAS for structural genomics
Bottlenecks and roadblocks in high-throughput XAS for structural genomics
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DOI：
10.1107/s0909049504028791
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发表时间：
2005-01-01
影响因子：
2.5
通讯作者：
Adams, MWW
中科院分区：
文献类型：
--
作者：
Scott, RA;Shokes, JE;Adams, MWW
生物体的整个蛋白质补体（蛋白质组）的结构和功能特征可以提供一个基础设施，在此基础上可以构建有关生物途径和系统生物学的问题。在许多结构基因组学和功能基因组学计划中，执行这种蛋白质组水平结构和功能表征所需的技术正在开发中。鉴于蛋白质组中金属活性位点的普遍存在，似乎有必要问一下，蛋白质组中金属位点的全面局部结构表征（金属蛋白质组学）是否是一个有效或可实现的目标。随着对所有金属蛋白活性位点结构的蛋白质组范围的了解，人们可以开始问金属插入、簇组装和金属蛋白表达如何受到生长条件或发育状态等的影响。高通量x射线吸收光谱（HTXAS）是一种研究金属蛋白质组的新技术。在建立从基因组到金属蛋白质组的管道时，必须克服金属位点结构高通量测定的几个瓶颈。例如，必须发明用于XAS检测的小样本阵列自动化，必须开发多个小体积低浓度样品快速数据采集的自动化，必须完善数据还原和分析的自动化。本文讨论了HTXAS开发的前景和缺陷，包括初步可行性实验的结果。
Structural and functional characterization of the entire protein complement (the proteome) of an organism can provide an infrastructure upon which questions about biological pathways and systems biology can be framed. The technology necessary to perform this proteome-level structural and functional characterization is under development in numerous structural genomics and functional genomics initiatives. Given the ubiquity of metal active sites in a proteome, it seems appropriate to ask whether comprehensive local structural characterization of metal sites within a proteome (metalloproteomics) is either a valid or obtainable goal. With a proteome-wide knowledge of the active-site structures of all metalloproteins, one could start to ask how metal insertion, cluster assembly and metalloprotein expression are affected by growth conditions or developmental status etc. High-throughput X-ray absorption spectroscopy (HTXAS) is being developed as a technology for investigating the metalloproteome. In creating a pipeline from genome to metalloproteome, several bottlenecks to high-throughput determination of metal-site structures must be overcome. For example, automation of arraying small samples for XAS examination must be invented, automation of rapid data collection of multiple low-volume low-concentration samples must be developed, automation of data reduction and analysis must be perfected. Discussed here are the promises and the pitfalls of HTXAS development, including the results of initial feasibility experiments.