RRC ID 50309
Author Tsutsui H, Jinno Y, Shoda K, Tomita A, Matsuda M, Yamashita E, Katayama H, Nakagawa A, Miyawaki A.
Title A diffraction-quality protein crystal processed as an autophagic cargo.
Journal Mol. Cell
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.
Volume 58(1)
Pages 186-93
Published 2015-4-2
DOI 10.1016/j.molcel.2015.02.007
PII S1097-2765(15)00099-4
PMID 25773597
MeSH Adaptor Proteins, Signal Transducing / genetics Adaptor Proteins, Signal Transducing / metabolism Animals Anthozoa / chemistry* Autophagy Crystallization Crystallography, X-Ray Cytosol / metabolism* Cytosol / ultrastructure Gene Expression Green Fluorescent Proteins / chemistry* Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism HEK293 Cells Hippocampus / cytology Hippocampus / metabolism Humans Lysosomes / metabolism* Lysosomes / ultrastructure Microtubule-Associated Proteins / genetics Microtubule-Associated Proteins / metabolism Models, Molecular Neurons / metabolism Neurons / ultrastructure Primary Cell Culture Protein Folding Protein Structure, Secondary Protein Transport Rats Recombinant Proteins / chemistry Recombinant Proteins / genetics Recombinant Proteins / metabolism Sequestosome-1 Protein X-Ray Diffraction
IF 14.248
Resource
DNA material Xpa (RDB15264) Xpa H62Q (RDB15265).