Antibody-Array Interaction Mapping, a New Method to Detect Protein Complexes Applied to the Discovery and Study of Serum Amyloid P Interactions with Kininogen in Human Plasma

作者：Bergsma Derek; Chen Songming; Buchweitz John; Gerszten Robert; Haab Brian B^*

来源：Molecular & Cellular Proteomics, 2010, 9(3): 446-456.

DOI：10.1074/mcp.M900418-MCP200

Protein-protein interactions are fundamentally important in biological processes, but the existing analytical tools have limited ability to sensitively and precisely measure the dynamic composition of protein complexes in biological samples. We report here the development of anti-body-array interaction mapping (AAIM) to address that need. We used AAIM to probe interactions among a set of 48 proteins in serum and found several known interactions as well potentially novel interactions, including multiprotein clusters of interactions. A novel interaction initially identified between the innate immune system protein C-reactive protein and the inflammatory protein kininogen (KNG) was confirmed in subsequent experiments to involve serum amyloid P instead of its highly related family member, C-reactive protein. AAIM was used in a variety of formats to further study this interaction. In vitro studies confirmed the ability of the purified proteins to interact and revealed a zinc dependence of the interaction. Studies using plasma samples collected longitudinally following a controlled myocardial infarction revealed no consistent changes in the serum amyloid P-KNG interaction levels but consistent changes in KNG activation and interactions with plasma prekallikrein. These results demonstrate a versatile platform for measuring the dynamic composition of protein complexes in biological samples that should have value for studies of normal and disease-related signaling networks, multiprotein clusters, or enzymatic cascades. Molecular & Cellular Proteomics 9: 446-456, 2010.

• 出版日期2010-3