Isolation, Characterization, and Imaging Analysis of Human Mediator Complexes Poster

Part # PS130

Abstract

¹Michael Ford, ¹Richard Jones, ¹Ravi Amunugama, ¹David Allen
¹MS Bioworks, 3950 Varsity Drive, Ann Arbor, MI, U.S.A. 48108
²Jacqui Mendez, ²Nancy Murphy, ²Marie Schwinn, ²Hélène Benink, ²Danette L. Daniels, and ²Marjeta Urh
²Promega Corporation, 2800 Woods Hollow Road, Madison, WI U.S.A. 53711

The understanding of protein complex assembly and mapping of protein interactions has rapidly grown in recent years due to significant advances in mass spectrometry. As experiments turn toward characterization of the human proteome and the complexity it presents, a need remains to efficiently capture complexes intact, particularly weak or transient interactors as well as large multiprotein complexes. Here we present a new technology based upon the use of a protein fusion tag, termed HaloTag, which allows for highly specific and covalent immobilization of proteins complexes, as well as the ability to do correlative cellular localization studies using fluorescent ligands. In studies presented here, this technology was used to further understanding of the multi-protein Mediator complex assemblies and their cellular function.

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