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Overview

TMPC is organized into research teams which address each of the constituent tasks of structural genomics. We also work closely with collaborators whose expertise in the function of membrane protein targets provides unique biological insight needed to advance our structure determination efforts and to guide interpretation and dissemination of results. Complementary cell-free and cell-based translation methods are currently being used to prepare proteins in the target classes of sphingolipid synthases, sigma-1 receptor, fatty acid desaturases, and pilin assembly complexes for functional studies and structure determination.

We are developing cell-free translation, which utilizes ENDEXT® technology, as a versatile method for studying membrane protein structure and function. TMPC’s experimental platform is based on establishing predictive behaviors for membrane proteins using free translation in small-scale (50 µL to 1.2 mL). Robotic translation has an advantage of simplicity and reproducibility in the provision of material used for further research and protocol development. When appropriate expression and solubilization conditions are identified, purification protocols are developed in small-scale, and then the work is transferred to different robots whose operational volume (1.2 mL to 20 mL) better supports structure determination or expanded biological studies.

The suite of cell-free translation robots in use at TMPC were developed by CellFree Sciences Co., Ltd. (Yokohama, Japan). These instruments, and the personnel and expertise to operate them, were assembled during PSI-1 and PSI-2. No other freely accessible site exists worldwide with this capability. We believe these robots will be a valuable resource for the PSI:Biology Network.

Other research infrastructure available at the TMPC and associated laboratories includes Sesame LIMS, Maxwell-16 Automated Purification System, AKTA Purification Systems, Shimadzu HPLC System with UV-Vis and fluorometric detection customized for use with detergents, TTP Lab Tech's Mosquito nanoliter crystallization robot, CrystalFarm Imaging System (Bruker AXS) with CrystalFarm Pro Data Management System, and routine access to GM/CA-CAT, SER-CAT, and LS-CAT at Advanced Photon Source.

Crystallization screening is being carried out in the TMPC facilities and at PSI:Biology Membrane Protein Structural Biology Consortium, led by Dr. Michael Malkowski.

 

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Annotation and Function

Cloning

Protein Expression and Purification

Reagents