Integrating in vivo crystallography and de novo protein design at the Nagoya University platform

In recent years, in vivo macromolecular crystallography (ivMX) has gained attention for studying protein crystals occurring naturally within cells and organisms. ivMX, also known as in cellulo MX, offers unique opportunities for high-throughput pipelines and workflow transformation by bypassing sample purification and crystallization steps. This eliminates the need for extensive protein preparation, as organisms perform these processes. Additionally, ivMX enables the study of proteins with post-translational modifications, which are often neglected in in vitro MX studies. 

Global research groups have made significant progress in understanding the unpredictable events governing natural crystal growth. Synchrotron SOLEIL in France has been crucial in initiating investigations into ivMX phenomena. Building upon this progress, Nagoya University has integrated the ivMX platform into the Next-BINDS national project, facilitating broader access and collaborative research, particularly in Japan.

This study presents the latest developments in the integrated ivMX platform, now open for project proposals. Our objectives include advancing the automation of the ivMX pipeline and streamlining sample handling, data collection, and analysis. We also aim to integrate de novo protein design techniques with ivMX, exploring protein structures beyond nature. By combining in vivo crystal growth and rational protein engineering, we anticipate breakthroughs in understanding structure-function relationships and novel applications in various scientific disciplines.

The integration of the ivMX platform at Nagoya University marks an exciting phase in in vivo protein crystal research. We invite researchers from diverse backgrounds to leverage this platform for their investigations. Together, we can unlock the full potential of ivMX, accelerating discoveries, and pioneering innovative approaches in structural biology and protein engineering.