Journal
JOURNAL OF PROTEOME RESEARCH
Volume 7, Issue 4, Pages 1712-1720Publisher
AMER CHEMICAL SOC
DOI: 10.1021/pr7007658
Keywords
protein interactions; Protein Interaction Reporter; PIR; electron transport; metal ion eduction; membrane proteins; FTICR-MS; outer membrane cytochrome C; OmcA; OmcB; MtrC; SO-0404
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Funding
- NCRR NIH HHS [R01 RR023334, S10 RR022538-01, R01 RR023334-01A1, 1 S10 RR017805-01, S10 RR022538] Funding Source: Medline
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Outer membrane (OM) cytochromes OmcA (SO1779) and MtrC (SO1778) are the integral components of electron transfer used by Shewanella oneidensis for anaerobic respiration of metal (hydr)oxides. Here the OmcA-MtrC interaction was identified in vivo using a novel hydrophobic chemical crosslinker (MRN) combined with immunoprecipitation techniques. In addition, identification of other OM proteins from the cross-linked complexes allows first visualization of the OmcA-MtrC interaction network. Further experiments on omcA and mtrC mutant cells showed OmcA plays a central role in the network interaction. For comparison, two commercial cross-linkers were also used in parallel, and both resulted in fewer OM protein identifications, indicating the superior properties of MRN for identification of membrane protein interactions. Finally, comparison experiments of in vivo cross-linking and cell lysate cross-linking resulted in significantly different protein interaction data, demonstrating the importance of in vivo cross-linking for study of protein-protein interactions in cells.
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