title: Protein microarrays for identification of novel extracellular protein-protein interactions.
abstract: Functional protein microarrays offer the capability for high-throughput protein interaction analysis and have long promised to be a powerful tool for understanding protein interactions at the proteome scale. Although popular techniques for protein-protein interaction mapping like yeast-two-hybrid and affinity-purification mass spectrometry have performed well for identifying intracellular protein-protein interactions, the study of interactions between extracellular proteins has remained challenging for these methods. Instead, the use of protein microarrays appears to be a robust and efficient method for the identification of interactions among the members of this class of protein. This unit describes methods for extracellular protein microarray production, screening, and analysis. A protocol is described for enhanced detection of low-affinity interactions by generating multivalent complexes using Fc-fusion bait proteins and protein A microbeads, along with a statistical method for hit scoring and identification of nonspecific interactions.
authors.affiliation: Department of Protein Chemistry, Genentech, South San Francisco, California, USA.
authors.forename: Irene
authors.initials: I
authors.lastname: Tom
authors.forename: Nicholas
authors.initials: N
authors.lastname: Lewin-Koh
authors.forename: Sree R
authors.initials: SR
authors.lastname: Ramani
authors.forename: Lino C
authors.initials: LC
authors.lastname: Gonzalez
mesh_terms: D000818:Animals; D005110:Extracellular Space / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000295:instrumentation* / Q000379:methods*; D011506:Proteins / Q000032:analysis* / Q000378:metabolism
chemical_list: D011506:Proteins