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

Molecular Cell
Hidekazu TsutsuiAtsushi Miyawaki

Abstract

Crystallization of proteins may occur in the cytosol of a living cell, but how a cell responds to intracellular protein crystallization remains unknown. We developed a variant of coral fluorescent protein that forms diffraction-quality crystals within mammalian cells. This expression system allowed the direct determination of its crystal structure at 2.9 Å, as well as observation of the crystallization process and cellular responses. The micron-sized crystal, which emerged rapidly, was a pure assembly of properly folded β-barrels and was recognized as an autophagic cargo that was transferred to lysosomes via a process involving p62 and LC3. Several lines of evidence indicated that autophagy was not required for crystal nucleation or growth. These findings demonstrate that in vivo protein crystals can provide an experimental model to study chemical catalysis. This knowledge may be beneficial for structural biology studies on normal and disease-related protein aggregation.

References

Feb 1, 1980·European Journal of Biochemistry·P M ColmanA Van Donkelaar
May 1, 1996·Microscopy Research and Technique·G Y FanL N Gastinel
Dec 2, 2000·Science·D J Klionsky, S D Emr
Jan 22, 2004·Biochemical and Biophysical Research Communications·Yuko HirotaYoshitaka Tanaka
May 1, 1997·Acta Crystallographica. Section D, Biological Crystallography·G N MurshudovE J Dodson
Feb 26, 2005·EMBO Reports·Hidekazu TsutsuiAtsushi Miyawaki
Oct 6, 2005·Physical Biology·Jonathan P K DoyeMichele Vendruscolo
Mar 3, 2007·Nature·Fasséli CoulibalyPeter Metcalf
Dec 25, 2007·Methods in Cell Biology·Satoshi Shimozono, Atsushi Miyawaki
Feb 8, 2008·Cell Structure and Function·Hiroyuki KatayamaAtsushi Miyawaki
Mar 7, 2009·Biochimica Et Biophysica Acta·Claudine KraftMatthias Peter
Jan 9, 2010·Acta Crystallographica. Section D, Biological Crystallography·Alexei Vagin, Alexei Teplyakov
Feb 16, 2010·FEBS Letters·Masaaki Komatsu, Yoshinobu Ichimura
Apr 13, 2010·Acta Crystallographica. Section D, Biological Crystallography·P EmsleyK Cowtan
Aug 27, 2011·Chemistry & Biology·Hiroyuki KatayamaAtsushi Miyawaki
Jan 31, 2012·Nature Methods·Rudolf KoopmannMichael Duszenko
Jan 1, 1997·Methods in Enzymology·Zbyszek Otwinowski, Wladek Minor

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Citations

Oct 7, 2015·Trends in Cell Biology·Aliaksandr KhaminetsIvan Dikic
Nov 27, 2015·Nature Communications·Yohendran BaskaranRobert C Robinson
Apr 7, 2016·Trends in Cell Biology·Nicholas T Ktistakis, Sharon A Tooze
Apr 2, 2016·Chemical Communications : Chem Comm·Satoshi AbeTakafumi Ueno
Jun 13, 2018·Biological Chemistry·Robert SchönherrLars Redecke
Apr 26, 2019·Journal of Biochemistry·Hideki NakamuraTakanari Inoue
Oct 31, 2019·Traffic·Celestin N MudogoChristian Betzel
Mar 7, 2020·Chemical Communications : Chem Comm·K MinamihataT Nagamune
Apr 7, 2016·Acta Crystallographica. Section D, Structural Biology·Marion BoudesFasséli Coulibaly
Dec 26, 2016·Histochemistry and Cell Biology·S NürnbergerD Barnewitz
Jul 15, 2017·International Journal of Molecular Sciences·Natalia V PovarovaKonstantin A Lukyanov
Jun 5, 2021·Nature Cell Biology·Dong-Jiunn Jeffery TruongGil Gregor Westmeyer
Sep 26, 2019·Nano Letters·Thomas L LiBianxiao Cui

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