Monti Maria, Orrù Stefania, Pagnozzi Daniela, Pucci Piero
CEINGE Biotecnologie Avanzate scarl and Dipartimento di Chimica Organica e Biochimica, Università di Napoli Federico II, via Comunale Margherita 482, Naples 80145, Italy.
Clin Chim Acta. 2005 Jul 24;357(2):140-50. doi: 10.1016/j.cccn.2005.03.019.
With the increase in the number of genome sequencing projects, there is a concomitant exponential growth in the number of protein sequences whose function is still unknown. Functional proteomics constitutes an emerging research area in the proteomic field whose approaches are addressed towards two major targets: the elucidation of the biological function of unknown proteins and the definition of cellular mechanisms at the molecular level.
The identification of interacting proteins in stable complexes in vivo is essentially achieved by affinity-based procedures. The basic idea is to express the protein of interest with a suitable tag to be used as a bait to fish its specific partners out from a cellular extract. Individual components within the multi-protein complex can then be identified by mass spectrometric methodologies.
The association of an unknown protein with partners belonging to a specific protein complex involved in a particular mechanism is strongly suggestive of the biological function of the protein. Moreover, the identification of protein partners interacting with a given protein will lead to the description of cellular mechanisms at the molecular level. The next goal will be to generate animal models bearing a tagged form of the bait protein.
随着基因组测序项目数量的增加,功能仍未知的蛋白质序列数量也随之呈指数增长。功能蛋白质组学是蛋白质组学领域中一个新兴的研究领域,其方法主要针对两个主要目标:阐明未知蛋白质的生物学功能以及在分子水平上定义细胞机制。
体内稳定复合物中相互作用蛋白质的鉴定主要通过基于亲和的方法来实现。基本思路是用合适的标签表达感兴趣的蛋白质,将其用作诱饵从细胞提取物中钓出其特定的伙伴。然后可以通过质谱方法鉴定多蛋白复合物中的各个组分。
未知蛋白质与参与特定机制的特定蛋白质复合物中的伙伴的关联强烈暗示了该蛋白质的生物学功能。此外,鉴定与给定蛋白质相互作用的蛋白质伙伴将有助于在分子水平上描述细胞机制。下一个目标将是生成携带诱饵蛋白质标签形式的动物模型。