Structural insight into substrate recognition by the endoplasmic reticulum folding-sensor enzyme: crystal structure of third thioredoxin-like domain of UDP-glucose:glycoprotein glucosyltransferase

BCL2-associated athanogene cochaperone 6 (Bag6) plays a central role in cellular homeostasis in a diverse array of processes and is part of the heterotrimeric Bag6 complex, which also includes ubiquitin-like 4A (Ubl4A) and transmembrane domain recognition complex 35 (TRC35). This complex recently has been shown to be important in the TRC pathway, the mislocalized protein degradation pathway, and the endoplasmic reticulum-associated degradation pathway. Here we define the architecture of the Bag6 complex, demonstrating that both TRC35 and Ubl4A have distinct C-terminal binding sites on Bag6 defining a minimal Bag6 complex. A crystal structure of the Bag6–Ubl4A dimer demonstrates that Bag6–BAG is not a canonical BAG domain, and this finding is substantiated biochemically. Remarkably, the minimal Bag6 complex defined here facilitates tail-anchored substrate transfer from small glutamine-rich tetratricopeptide repeat-containing protein α to TRC40. These findings provide structural insight into the complex network of proteins coordinated by Bag6.

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Crystal Structure of the Epo1-Bem3 Complex for Bud Growth

International Journal of Molecular Sciences ◽

10.3390/ijms22083812 ◽

2021 ◽

Vol 22 (8) ◽

pp. 3812

Author(s):

Jin Wang ◽

Lei Li ◽

Zhenhua Ming ◽

Lijie Wu ◽

Liming Yan

Keyword(s):

Crystal Structure ◽

Endoplasmic Reticulum ◽

Membrane Protein ◽

Protein Complex ◽

Molecular Architecture ◽

Hydrogen Deuterium ◽

Bud Growth ◽

Structural Insight ◽

Guanosine Triphosphatase ◽

Insight Into

Tubules of the endoplasmic reticulum (ER) spread into the buds of yeast by an actin-based mechanism and, upon entry, become attached to the polarisome, a proteinaceous micro-compartment below the tip of the bud. The minimal tether between polarisome and cortical ER is formed by a protein complex consisting of Epo1, a member of the polarisome, Scs2, a membrane protein of the ER and Cdc42 guanosine triphosphatase-activating protein Bem3. Here, we report the crystal structure of a complex between Epo1 and Bem3. In addition, we characterize through the hydrogen/deuterium (H/D) exchange assay the interface between Scs2 and Epo1. Our findings provide a first structural insight into the molecular architecture of the link between cortical ER and the polarisome.

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Crystal structure of the Ube2K/E2-25K and K48-linked di-ubiquitin complex provides structural insight into the mechanism of K48-specific ubiquitin chain synthesis

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2018.10.067 ◽

2018 ◽

Vol 506 (1) ◽

pp. 102-107 ◽

Cited By ~ 2

Author(s):

Jung-Gyu Lee ◽

Hyung-Seop Youn ◽

Jung Youn Kang ◽

Sam-Yong Park ◽

Akinori Kidera ◽

...

Keyword(s):

Crystal Structure ◽

Ubiquitin Chain ◽

Structural Insight ◽

Chain Synthesis ◽

Insight Into

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Structural Insight Into Protein Binding of Boron Tracedrug UTX-97 Revealed by the Co-Crystal Structure With Lysozyme at 1.26 Å Resolution

Journal of Pharmaceutical Sciences ◽

10.1016/j.xphs.2016.06.005 ◽

2016 ◽

Vol 105 (8) ◽

pp. 2298-2301

Author(s):

Yukio Morimoto ◽

Hideko Nagasawa ◽

Yoshihiro Uto ◽

Toshiyuki Chatake ◽

Hitoshi Hori

Keyword(s):

Crystal Structure ◽

Protein Binding ◽

Structural Insight ◽

Insight Into

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Structural Insight into the Dioxygenation of Nitroarene Compounds: the Crystal Structure of Nitrobenzene Dioxygenase

Journal of Molecular Biology ◽

10.1016/j.jmb.2005.03.052 ◽

2005 ◽

Vol 348 (5) ◽

pp. 1139-1151 ◽

Cited By ~ 72

Author(s):

Rosmarie Friemann ◽

Maja M. Ivkovic-Jensen ◽

Daniel J. Lessner ◽

Chi-Li Yu ◽

David T. Gibson ◽

...

Keyword(s):

Crystal Structure ◽

Structural Insight ◽

Insight Into

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Crystal structure of Gig2 protein from Candida albicans provides a structural insight into DUF1479 family oxygenases

International Journal of Biological Macromolecules ◽

10.1016/j.ijbiomac.2019.10.138 ◽

2020 ◽

Vol 150 ◽

pp. 1272-1280

Author(s):

Priya Rani ◽

Gunjan Gautam ◽

Tamanna Anwar ◽

Samudrala Gourinath ◽

Asis Datta

Keyword(s):

Crystal Structure ◽

Candida Albicans ◽

Structural Insight ◽

Insight Into

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Structural Insight into the Antibiotic Action of Telithromycin against Resistant Mutants

Journal of Bacteriology ◽

10.1128/jb.185.14.4276-4279.2003 ◽

2003 ◽

Vol 185 (14) ◽

pp. 4276-4279 ◽

Cited By ~ 131

Author(s):

Rita Berisio ◽

Joerg Harms ◽

Frank Schluenzen ◽

Raz Zarivach ◽

Harly A. S. Hansen ◽

...

Keyword(s):

Crystal Structure ◽

Deinococcus Radiodurans ◽

Ribosomal Subunit ◽

Large Ribosomal Subunit ◽

Resistant Strains ◽

Structural Insight ◽

Resistant Mutants ◽

Exit Tunnel ◽

Structural Insights ◽

Insight Into

ABSTRACT The crystal structure of the ketolide telithromycin bound to the Deinococcus radiodurans large ribosomal subunit shows that telithromycin blocks the ribosomal exit tunnel and interacts with domains II and V of the 23S RNA. Comparisons to other clinically relevant macrolides provided structural insights into its enhanced activity against macrolide-resistant strains.

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Structural studies on dehydroshikimate dehydratase

Acta Crystallographica Section A Foundations and Advances ◽

10.1107/s2053273314095242 ◽

2014 ◽

Vol 70 (a1) ◽

pp. C475-C475

Author(s):

James Peek ◽

Dinesh Christendat

Keyword(s):

Sole Carbon Source ◽

Catalytic Mechanism ◽

Substrate Recognition ◽

Structural Studies ◽

Sugar Phosphate ◽

X Ray ◽

Structural Insight ◽

Functional Relevance ◽

Insight Into

The soil bacterium, Pseudomonas putida, is capable of using the alicyclic compound quinate as a sole carbon source. During this process, quinate is converted to 3-dehydroshikimate, which subsequently undergoes a dehydration to form protocatechuate. The latter transformation is performed by the enzyme dehydroshikimate dehydratase (DSD). We have recombinantly produced DSD from P. putida and are currently performing x-ray crystallographic studies on the enzyme to gain structural insight into its catalytic mechanism and mode of substrate recognition. Initial crystals of DSD diffracted to 2.7 Ä resolution, but exhibited strong twinning. A redesigned construct has recently yielded crystals that diffract to similar resolution, but with a significantly reduced tendency toward twinning. Interestingly, sequence analysis of P. putida DSD reveals that the protein is in fact a fusion of two distinct domains: an N-terminal sugar phosphate isomerase-like domain associated with DSD activity, and a C-terminal hydroxyphenylpyruvate dioxygenase (HPPD)-like domain with unknown functional significance. Structural characterization of the protein may provide novel insight into the functional relevance of the unusual HPPD-like domain.

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