Manjasetty Babu A, Turnbull Andrew P, Panjikar Santosh, Büssow Konrad, Chance Mark R
Case Center for Synchrotron Biosciences, National Synchrotron Light Source, Brookhaven National Laboratory, Upton, NY11973, USA.
Proteomics. 2008 Feb;8(4):612-25. doi: 10.1002/pmic.200700687.
The sequence infrastructure that has arisen through large-scale genomic projects dedicated to protein analysis, has provided a wealth of information and brought together scientists and institutions from all over the world. As a consequence, the development of novel technologies and methodologies in proteomics research is helping to unravel the biochemical and physiological mechanisms of complex multivariate diseases at both a functional and molecular level. In the late sixties, when X-ray crystallography had just been established, the idea of determining protein structure on an almost universal basis was akin to an impossible dream or a miracle. Yet only forty years after, automated protein structure determination platforms have been established. The widespread use of robotics in protein crystallography has had a huge impact at every stage of the pipeline from protein cloning, over-expression, purification, crystallization, data collection, structure solution, refinement, validation and data management- all of which have become more or less automated with minimal human intervention necessary. Here, recent advances in protein crystal structure analysis in the context of structural genomics will be discussed. In addition, this review aims to give an overview of recent developments in high throughput instrumentation, and technologies and strategies to accelerate protein structure/function analysis.
通过致力于蛋白质分析的大规模基因组计划所形成的序列基础设施,提供了丰富的信息,并汇聚了来自世界各地的科学家和机构。因此,蛋白质组学研究中新技术和方法的发展有助于在功能和分子水平上揭示复杂多变量疾病的生化和生理机制。在六十年代后期,当X射线晶体学刚刚建立时,在几乎普遍的基础上确定蛋白质结构的想法就像是一个不可能实现的梦想或奇迹。然而,仅仅四十年后,就建立了自动化的蛋白质结构测定平台。机器人技术在蛋白质晶体学中的广泛应用在从蛋白质克隆、过量表达、纯化、结晶、数据收集、结构解析、精修、验证到数据管理的整个流程的每个阶段都产生了巨大影响——所有这些流程或多或少都已实现自动化,只需最少的人工干预。在此,将讨论结构基因组学背景下蛋白质晶体结构分析的最新进展。此外,本综述旨在概述高通量仪器以及加速蛋白质结构/功能分析的技术和策略的最新发展。