Interactome Mapping: Using Protein Microarray Technology to Reconstruct Diverse Protein Networks

Uzoma, Ijeoma; Zhu, Heng

In 2001 the Snyder group reported the fabrication of the first proteome microarray in the budding yeast, representing a major advance for the field [12] . In order to construct this array, approximately 5800 full-length yeast ORFs were individually expressed in yeast and their protein products purified as N-terminal GST-fusion proteins. Each purified protein was then robotically spotted on a single glass slide in duplicate at high density to form the first ''proteome'' microarray, covering more than 75% of the yeast proteome. More recently, proteome microarrays have been fabricated from the proteomes of viruses, bacteria, plants and humans [8, [13] [14] [15] [16] . Functional protein microarrays have been successfully applied to identify protein-protein, protein-lipid, protein-antibody, protein-small molecules, protein-DNA, protein-RNA, protein-lectin and lectin-cell interactions [8, 9, 12, 14, [16] [17] [18] [19] , to identify substrates or enzymes for phosphorylation, ubiquitylation, acetylation and nitrosylation [11, [20] [21] [22] [23] [24] , as well as to profile immune response [25] . In this review, we will focus on inventive applications for protein microarrays and the significant findings that contribute to understanding the complex interactomes within cells ( Table 1) .