The structure of the PA28–20S proteasome complex from Plasmodium falciparum and implications for proteostasis

Xie, Stanley C.; Metcalfe, Riley D.; Hanssen, Eric; Yang, Tuo; Gillett, David L.; Leis, Andrew; Morton, Craig J.; Kuiper, Michael J.; Parker, Michael W.; Spillman, Natalie J.; Wong, Wilson; Tsu, Christopher; Dick, Lawrence R.; Griffin, Michael D. W.; Tilley, Leann

doi:10.1038/s41564-019-0524-4

Cited by 35 publications

(62 citation statements)

References 86 publications

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“…In a recent study, PA28 from Plasmodium falciparum (PfPA28) was found to exhibit a heptameric structure with a charge-segregated pore formed by an approximately 20-Å positively charged apical (narrow) end and a 35-Å negatively charged basal end. The size of the pore (20 Å minimum) is sufficient for peptides and unfolded polypeptide substrates/products to pass through [14]. Given that the size of the two charged openings of PA200 (19.6 × 13.7 Å and 23.3 × 17.8 Å) in our study is similar to the pores of PfPA28 (Fig 5D) [14], it seems plausible that the two openings of PA200 are the gates for feeding substrates for proteolysis.…”

Section: Plos Biologymentioning

confidence: 59%

“…The size of the pore (20 Å minimum) is sufficient for peptides and unfolded polypeptide substrates/products to pass through [14]. Given that the size of the two charged openings of PA200 (19.6 × 13.7 Å and 23.3 × 17.8 Å) in our study is similar to the pores of PfPA28 (Fig 5D) [14], it seems plausible that the two openings of PA200 are the gates for feeding substrates for proteolysis. Intriguingly, the positively charged residues of these two openings respectively bind 5,6[PP]2-InsP 4 and InsP 6 (Fig 5D, S4 Fig), which was confirmed by our HPLC-MS results (Fig 6).…”

Section: Plos Biologymentioning

confidence: 59%

“…PA200 makes numerous contacts with the α-ring surface of 20S by using the C-terminal hydrophobic-tyrosine-other (HbYX) motif and a loop (T562 to K574) to contact the N-terminals of 20S subunits α5, α6 and α1, α2 (Fig 4A-4F). The HbYX motif of PA200 (YYA) inserts into the pocket between proteasome subunits α5 and α6 in a manner similar to Blm10, 11S activator (PA26 and PA28), and 19S/PAN activators [14,21]. These interactions play a crucial role in displacing the Pro17 turn and subsequent opening of the proteasome gate.…”

Section: Plos Biologymentioning

confidence: 99%

“…This factor is an ATP-dependent activator that binds and unfolds ubiquitin-conjugated proteins to promote proteasomal degradation of its substrates [11]. The other three activator families-i.e., the 11S (PA28α/β), PA28γ (also referred to as REGγ), and PA200-are ATP-independent and less broadly conserved than the ATP-dependent activators [2,[12][13][14]. Typically, proteasomes in mammalian cells come in two predominant forms: the activator-unbound 20S proteasomes (CP) and 26S holoenzymes (CP-19S complex).…”

Section: Introductionmentioning

confidence: 99%

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Cryo-EM structures of the human PA200 and PA200-20S complex reveal regulation of proteasome gate opening and two PA200 apertures

Guan

Wang

et al. 2020

PLoS Biol

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Proteasomes are highly abundant and conserved protease complexes that eliminate unwanted proteins in the cells. As a single-chain ATP-independent nuclear proteasome activator, proteasome activator 200 (PA200) associates with 20S core particle to form proteasome complex that catalyzes polyubiquitin-independent degradation of acetylated histones, thus playing a pivotal role in DNA repair and spermatogenesis. Here, we present cryo-electron microscopy (cryo-EM

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Section: Plos Biologymentioning

confidence: 59%

Section: Plos Biologymentioning

confidence: 59%

Section: Plos Biologymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Cryo-EM structures of the human PA200 and PA200-20S complex reveal regulation of proteasome gate opening and two PA200 apertures

Guan

Wang

et al. 2020

PLoS Biol

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“…The proteasome has been intensely studied from a structural and functional point of view since its discovery in 1988 15 . Electron microscopy (EM) allowed to observe the 20S in complex with different regulators [16][17][18][19][20][21] together with certain catalytic intermediate-states of the 26S 22 . Covalent cross-linking coupled to mass spectrometry (MS) helped refining 26S structures 23,24 and generating structural models involving different partners, including the 19S 18 , Ecm29 11 and other PIPs 11 .…”

Section: Main Text Introductionmentioning

confidence: 99%

Conformational maps of human 20S proteasomes reveal PA28- and immuno-dependent inter-ring crosstalks

Lesné

Locard‐Paulet

Parra

et al. 2020

Preprint

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Here, we used for the first time Hydrogen-Deuterium eXchange coupled to Mass Spectrometry (HDX-MS) to investigate conformational differences between the human standard 20S (std20S) and immuno 20S (i20s) proteasomes alone or in complex with PA28αβ or PA28γ activators. Their solvent accessibility was analyzed through a dedicated bioinformatic pipeline including stringent statistical analysis and 3D visualization. These data confirmed the existence of allosteric differences between the std20S and i20S at the surface of the α-ring triggered from inside the catalytic β-ring. Additionally, binding of the PA28 regulators to the 20S proteasomes modified solvent accessibility due to conformational changes of the β-rings. This work is not only a proof-of-concept that HDX-MS can be used to get structural insights on large multi-protein complexes in solution, it also demonstrates that the binding of the std20S or i20S subtype to any of its PA28 activator triggers allosteric changes that are specific to this 20S/PA28 pair.

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Development of Potent and Highly Selective Epoxyketone‐Based Plasmodium Proteasome Inhibitors

Almaliti

Fajtová

Calla

et al. 2023

Chemistry A European J

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Here we present remarkable epoxyketone-based proteasome inhibitors with low nanomolar in vitro potency for bloodstage Plasmodium falciparum and low cytotoxicity for human cells. Our best compound has more than 2,000-fold greater selectivity for erythrocytic-stage P. falciparum over HepG2 and H460 cells, which is largely driven by the accommodation of the parasite proteasome for a D-amino acid in the P3 position and the preference for a difluorobenzyl group in the P1 position. We isolated the proteasome from P. falciparum cell extracts and determined that the best compound is 171-fold more potent at inhibiting the β5 subunit of P. falciparum proteasome when compared to the same subunit of the human constitutive proteasome. These compounds also significantly reduce parasitemia in a P. berghei mouse infection model and prolong survival of animals by an average of 6 days. The current epoxyketone inhibitors are ideal starting compounds for orally bioavailable antimalarial drugs.

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The structure of the PA28–20S proteasome complex from Plasmodium falciparum and implications for proteostasis

Cited by 35 publications

References 86 publications

Cryo-EM structures of the human PA200 and PA200-20S complex reveal regulation of proteasome gate opening and two PA200 apertures

Cryo-EM structures of the human PA200 and PA200-20S complex reveal regulation of proteasome gate opening and two PA200 apertures

Conformational maps of human 20S proteasomes reveal PA28- and immuno-dependent inter-ring crosstalks

Development of Potent and Highly Selective Epoxyketone‐Based Plasmodium Proteasome Inhibitors

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