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Related Experiment Video

Updated: Apr 16, 2026

$Microcrystallography of Protein Crystals and In Cellulo Diffraction$

Microcrystallography of Protein Crystals and In Cellulo Diffraction

Published on: July 21, 2017

A diffraction-quality protein crystal processed as an autophagic cargo.

Hidekazu Tsutsui¹, Yuka Jinno², Keiko Shoda³

¹Laboratory for Cell Function Dynamics, Brain Science Institute, RIKEN, Wako, Saitama 351-0198, Japan; Department of Material Science, Japan Advanced Institute of Science and Technology, Nomi, Ishikawa 923-1292, Japan; PRESTO, Japan Science and Technology Agency (JST), 4-1-8 Hon-cho, Kawaguchi, Saitama 351-0198, Japan.

|March 17, 2015

Summary

Cells can respond to intracellular protein crystallization by engulfing protein crystals via autophagy. This process involves p62 and LC3, but autophagy is not essential for crystal formation, offering insights into protein aggregation.

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Published on: March 28, 2011

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Last Updated: Apr 16, 2026

$Microcrystallography of Protein Crystals and In Cellulo Diffraction$

Microcrystallography of Protein Crystals and In Cellulo Diffraction

Published on: July 21, 2017

Use of a Robot for High-throughput Crystallization of Membrane Proteins in Lipidic Mesophases

Use of a Robot for High-throughput Crystallization of Membrane Proteins in Lipidic Mesophases

Published on: September 1, 2012

Crystallization of Membrane Proteins in Lipidic Mesophases

Crystallization of Membrane Proteins in Lipidic Mesophases

Published on: March 28, 2011

Area of Science:

Cell biology
Structural biology
Biochemistry

Background:

Intracellular protein crystallization can occur, but cellular responses are not well understood.
Studying in vivo protein crystallization is challenging due to limitations in experimental models.

Purpose of the Study:

To develop an experimental system for observing intracellular protein crystallization and cellular responses in real-time.
To determine the crystal structure of an intracellular protein crystal and elucidate the cellular mechanisms involved in its clearance.

Main Methods:

Engineered a coral fluorescent protein variant for intracellular crystallization in mammalian cells.
Determined the crystal structure using X-ray diffraction.
Observed crystallization dynamics and cellular responses using microscopy.
Investigated the role of autophagy in crystal clearance through genetic and biochemical approaches.

Main Results:

Successfully generated diffraction-quality protein crystals within mammalian cells.
Determined the 2.9 Å crystal structure of the coral fluorescent protein variant.
Observed rapid formation of micron-sized, pure β-barrel crystals.
Demonstrated that protein crystals are recognized as autophagic cargo and trafficked to lysosomes via p62 and LC3.
Showed that autophagy is not required for crystal nucleation or growth.

Conclusions:

In vivo protein crystals can serve as a model system for studying chemical catalysis.
Autophagy plays a role in clearing intracellular protein crystals, independent of their formation.
Findings advance understanding of cellular responses to protein aggregation and have implications for structural biology.