On the Trails of the Proteasome Fold: Structural and Functional Analysis of the Ancestral β-Subunit Protein Anbu

Vielberg, M.-T.; Bauer, Verena C.; Groll, M.

doi:10.1016/j.jmb.2018.01.004

Cited by 4 publications

(6 citation statements)

References 49 publications

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“…Thus, it is suggested that Anbu represents the common origin of proteasome subunits. Based on the Anbu crystal structure (PDB ID: 5NYP), the amber codon was placed at the solvent‐exposed position Asp71. After expression of Anbu‐71TAG in the presence of 3 and the BioRS/tRNA CUA pair, the cell lysate was equilibrated with basic buffer (pH 11) prior to purification.…”

Section: Figurementioning

confidence: 99%

Genetically Encoded Biotin Analogues: Incorporation and Application in Bacterial and Mammalian Cells

et al. 2019

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The biotin–streptavidin interaction is among the strongest known in nature. Herein, the site‐directed incorporation of biotin and 2‐iminobiotin composed of noncanonical amino acids (ncAAs) into proteins is reported. 2‐Iminobiotin lysine was employed for protein purification based on the pH‐dependent dissociation constant to streptavidin. By using the high‐affinity binding of biotin lysine, the bacterial protein RecA could be specifically isolated and its interaction partners analyzed. Furthermore, the biotinylation approach was successfully transferred to mammalian cells. Stringent control over the biotinylation site and the tunable affinity between ncAAs and streptavidin of the different biotin analogues make this approach an attractive tool for protein interaction studies, protein immobilization, and the generation of well‐defined protein–drug conjugates.

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Section: Figurementioning

confidence: 99%

Genetically Encoded Biotin Analogues: Incorporation and Application in Bacterial and Mammalian Cells

et al. 2019

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“…Interestingly, Anbu is found in different oligomeric states in different organisms. So far, both dodecameric assemblies consisting of six and tetradecameric assemblies consisting of seven dimers have been observed, which underlines the apparent evolutionary ease of oligomerization transitions in absence of dihedral symmetry constraints. One key element of the helical assembly is a coiled coil that forms between the long C‐terminal α helices of the opposing protomers, spanning the dimer interface and stabilizing the dimer significantly .…”

Section: Dimerization As a First Step Towards Self‐compartmentalizationmentioning

confidence: 99%

“…The recent identification and characterization of two further bacterial self‐compartmentalizing NTN‐hydrolases, ancestral β subunit (Anbu), and betaproteobacterial proteasome homolog (BPH), added more diversity to this family (Figure ). While BPH was found to form tetradecamers of D7 symmetry similar to the D6‐symmetric HslV, Anbu revealed an unexpected helical oligomerization architecture of two stacked open rings without dihedral symmetry, resembling a split lock washer with C2 symmetry . The evolutionary history of the proteasome family, especially of the 20S proteasome and HslV, has long been controversial.…”

Section: Introductionmentioning

confidence: 99%

On the Origins of Symmetry and Modularity in the Proteasome Family

Fuchs

Hartmann

2019

BioEssays

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The proteasome family of proteases comprises oligomeric assemblies of very different symmetry. In different sizes, it features ring‐like oligomers with dihedral symmetry that allow the stacking of further rings of regulatory subunits as observed in the modular proteasome system, but also less symmetric helical assemblies. Comprehensive sequence and structural analyses of proteasome homologs reveal a parsimonious scenario of how symmetry may have emerged from a monomeric ancestral precursor and how it may have evolved throughout the proteasome family. The four characterized representatives—ancestral β subunit (Anbu), HslV, betaproteobacterial proteasome homolog (BPH), and the 20S proteasome—are outlasting cornerstones in the family's evolutionary history, each marking a transition in symmetry. This article contextualizes the evolutionary and functional key aspects of these symmetry transitions, explaining how they facilitated the diversification and concurrent evolution of independent proteolytic systems side by side, each with its customized network of auxiliary interactors.

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“…While heat shock locus V (HslV) ( Rohrwild et al , 1996 ), which is found in bacteria and some eukaryotic organelles of endosymbiotic origin, was the only known proteasome homolog for many years, we and others have recently characterized three further proteasome homologs: the bacterial complexes Anbu ( Fuchs et al , 2017 ; Gille et al , 2003 ; Piasecka et al , 2018 ; Valas and Bourne, 2008 ; Vielberg et al , 2018 ) and BPH ( Fuchs et al , 2018a ), and the monomeric archaeal connectase ( Fuchs et al , 2021 ). In the Pfam database ( Finn et al , 2014 ), the proteasome subunits, HslV, BPH and Anbu are classified into the ‘Proteasome subunit’ family within the ‘NTN hydrolase’ clan (N-terminal nucleophile).…”

Section: Introductionmentioning

confidence: 99%

“…We have previously proposed that BPH may instead have functions similar to those of the uncapped proteasome ( Fuchs et al , 2018a ). Likewise, no function is known for the Anbu complex, but its unique split-ring architecture suggests a different mode of action ( Fuchs et al , 2017 ; Piasecka et al , 2018 ; Vielberg et al , 2018 ). Lastly, unlike the other proteasome-like homologs, connectase is not a protease, but instead acts as a highly specific protein ligase ( Fuchs et al , 2021 ).…”

Section: Introductionmentioning

confidence: 99%

An astonishing wealth of new proteasome homologs

2021

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Motivation The proteasome is the main proteolytic machine for targeted protein degradation in archaea and eukaryotes. While some bacteria also possess the proteasome, most of them contain a simpler and more specialized homolog, the HslV protease. In recent years, three further homologs of the proteasome core subunits have been characterized in prokaryotes: Anbu, BPH, and connectase. With the inclusion of these members, the family of proteasome-like proteins now exhibits a range of architectural and functional forms, from the canonical proteasome, a barrel-shaped protease without pronounced intrinsic substrate specificity, to the monomeric connectase, a highly specific protein ligase. Results We employed systematic sequence searches to show that we have only seen the tip of the iceberg so far and that beyond the hitherto known proteasome homologs lies a wealth of distantly related, uncharacterized homologs. We describe a total of 22 novel proteasome homologs in bacteria and archaea. Using sequence and structure analysis, we analyze their evolutionary history and assess structural differences that may modulate their function. With this initial description, we aim to stimulate the experimental investigation of these novel proteasome-like family members. Availability The protein sequences in this study are searchable in the MPI Bioinformatics Toolkit (https://toolkit.tuebingen.mpg.de) with ProtBLAST/PSI-BLAST and with HHpred (database "proteasome_homologs"). The following data are available at https://data.mendeley.com/datasets/t48yhff7hs/3: (I) sequence alignments for each proteasome-like homolog, (II) the coordinates for their structural models, and (III) a cluster-map file, which can be navigated interactively in CLANS and gives direct access to all the sequences in this study. Supplementary information Supplementary data are available at Bioinformatics online.

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On the Trails of the Proteasome Fold: Structural and Functional Analysis of the Ancestral β-Subunit Protein Anbu

Cited by 4 publications

References 49 publications

Genetically Encoded Biotin Analogues: Incorporation and Application in Bacterial and Mammalian Cells

Genetically Encoded Biotin Analogues: Incorporation and Application in Bacterial and Mammalian Cells

On the Origins of Symmetry and Modularity in the Proteasome Family

An astonishing wealth of new proteasome homologs

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