In this project, MicroFab will develop a high throughput ink-jet based system to screen for the initial signs of crystallization of membrane proteins, while maximizing the conservation of limited protein samples. Investigators can then repeat successful crystallization experiments determined from the initial screens to grow larger well-ordered crystals for diffraction experiments. The structure of membrane proteins obtained by x-ray crystallography can provide information for computational and structure based drug development. Unfortunately, the crystallization of integral membrane proteins often fails to yield crystals useful for diffraction analysis. Membrane protein (MP) crystallization difficulties can be attributed to their very narrow metastable solubility regions and scarcity of material. The consensus solution to these problems is to decrease protein consumption and narrow the sampling of crystal growth conditions by using smaller screening volumes. Ink-jet dispensing is a technology capable of reliably delivering small volumes ranging from picoliter to microliter, the later through accretion. In Phase I MicroFab will determine the operational parameters required to dispense protein and detergent screening solutions using ink-jet devices. MicroFab will also determine the optimum process for coalescing and mixing of these solutions under oil. Finally, MicroFab will conduct initial crystallization condition screens of membrane proteins using ink-jet dispensing. We expect these experiments to provide critical information that will ultimately enable high throughput picoliter MP crystal growth assays as a mainstream technique. ? ?
