DNA-mediated proteolysis by neutrophil elastase enhances binding activities of the HMGB1 protein

Wang, Xi; Mayorga-Flores, Marlen; Bien, Karina G.; Bailey, Aaron O.; Iwahara, Junji

doi:10.1016/j.jbc.2022.102577

Cited by 8 publications

(9 citation statements)

References 71 publications

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“…Three successive steps have been previously proposed for HMGB1 binding to the TLR4/MD2 complex ( 59 ): 1) the A-box first binds to TLR4 through R24, K28 and the tip of the first helix of HMGB1; 2) then the B-box can bind MD2; 3) The B-box then mediates TLR4 dimerization and signaling. (B) The lack of the AcTail in the F1 fragment likely enhances binding to DNA and to the TLR4/MD2 complex, as shown for other fragments similarly lacking the AcTail ( 60 ). In this case, LPS may not be necessary for binding.…”

Section: Discussionmentioning

confidence: 87%

“…This is true for charged ligands such as DNA, because intramolecular interactions of the DE repeats with the DNA-binding A- or B-boxes and their intermolecular interaction with DNA are mutually exclusive ( 58 , 61 ). In the same line, removal of the C-terminal acidic tail has been shown to enhance HMGB1 binding affinity for the TLR4/MD-2 complex in absence of LPS in vitro ( 60 ), while retaining similar TNF inflammatory signaling properties ( 27 ). The same hypothesis may apply to TLR2 binding ( 62 ).…”

Section: Discussionmentioning

confidence: 98%

“…For example, when the second nuclear localization signal of HMGB1 (NLS2, residues 179-185) is missing, the way back to the nucleus is closed ( 3 ). Because the acidic DE repeats in the acidic tail are involved in dynamic autoinhibition of HMGB1 in the free state, their removal promotes some binding activities of HMGB1 ( 60 ). This is true for charged ligands such as DNA, because intramolecular interactions of the DE repeats with the DNA-binding A- or B-boxes and their intermolecular interaction with DNA are mutually exclusive ( 58 , 61 ).…”

Section: Discussionmentioning

confidence: 99%

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HMGB1 cleavage by complement C1s and its potent anti-inflammatory product

et al. 2023

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Complement C1s association with the pathogenesis of several diseases cannot be simply explained only by considering its main role in activating the classical complement pathway. This suggests that non-canonical functions are to be deciphered for this protease. Here the focus is on C1s cleavage of HMGB1 as an auxiliary target. HMGB1 is a chromatin non-histone nuclear protein, which exerts in fact multiple functions depending on its location and its post-translational modifications. In the extracellular compartment, HMGB1 can amplify immune and inflammatory responses to danger associated molecular patterns, in health and disease. Among possible regulatory mechanisms, proteolytic processing could be highly relevant for HMGB1 functional modulation. The unique properties of HMGB1 cleavage by C1s are analyzed in details. For example, C1s cannot cleave the HMGB1 A-box fragment, which has been described in the literature as an inhibitor/antagonist of HMGB1. By mass spectrometry, C1s cleavage was experimentally identified to occur after lysine on position 65, 128 and 172 in HMGB1. Compared to previously identified C1s cleavage sites, the ones identified here are uncommon, and their analysis suggests that local conformational changes are required before cleavage at certain positions. This is in line with the observation that HMGB1 cleavage by C1s is far slower when compared to human neutrophil elastase. Recombinant expression of cleavage fragments and site-directed mutagenesis were used to confirm these results and to explore how the output of C1s cleavage on HMGB1 is finely modulated by the molecular environment. Furthermore, knowing the antagonist effect of the isolated recombinant A-box subdomain in several pathophysiological contexts, we wondered if C1s cleavage could generate natural antagonist fragments. As a functional readout, IL-6 secretion following moderate LPS activation of RAW264.7 macrophage was investigated, using LPS alone or in complex with HMGB1 or some recombinant fragments. This study revealed that a N-terminal fragment released by C1s cleavage bears stronger antagonist properties as compared to the A-box, which was not expected. We discuss how this fragment could provide a potent brake for the inflammatory process, opening the way to dampen inflammation.

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Section: Discussionmentioning

confidence: 87%

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

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HMGB1 cleavage by complement C1s and its potent anti-inflammatory product

et al. 2023

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“…Due to the cations condensed around the negatively charged macromolecule, the total Na + concentration in the solution is higher than the Na + concentration in the original buffer, and the slope in the plot corresponds to the number of condensed cations ΔN cation . 21 base pairs) named J1, 26 which exhibits high affinities for the full-length HMGB1 8,27 and the Δ30 variant (the binding affinity data are shown in Figure S9). When unlabeled J1 was added to a solution of the complex of 15 N DERT30 and the unlabeled Δ30 variant, the signals from 15 N-labeled DERT30 moved toward the chemical shifts for the free state (Figure 4A, middle).…”

mentioning

confidence: 99%

“…Hundreds of human proteins have low-complexity sequences of consecutive aspartate (D) or glutamate (E) residues, which are referred to as D/E repeats (examples shown in Figure A). , Approximately 50% of proteins containing D/E repeats are DNA/RNA-binding proteins. For some of them, the D/E repeats interact with the DNA-binding domains within the same polypeptide chain and diminish the association with DNA, causing autoinhibition. − Other D/E repeats can exhibit chaperone-like activity. , Currently, however, very little is known about the physicochemical properties of the D/E repeats. In this article, using NMR spectroscopy, we demonstrate that D/E repeats have electrostatic properties similar to those of DNA and electrostatically influence the DNA-binding domains via intramolecular interactions, competing with DNA.…”

mentioning

confidence: 99%

Influence of an Intrinsically Disordered Region on Protein Domains Revealed by NMR-Based Electrostatic Potential Measurements

Yu,

Wang,

Tan

et al. 2024

J. Am. Chem. Soc.

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Many human proteins possess intrinsically disordered regions containing consecutive aspartate or glutamate residues (“D/E repeats”). Approximately half of them are DNA/RNA-binding proteins. In this study, using nuclear magnetic resonance (NMR) spectroscopy, we investigated the electrostatic properties of D/E repeats and their influence on folded domains within the same protein. Local electrostatic potentials were directly measured for the HMGB1 protein, its isolated D/E repeats, and DNA-binding domains by NMR. The data provide quantitative information about the electrostatic interactions between distinct segments of HMGB1. Due to the interactions between the D/E repeats and the DNA-binding domains, local electrostatic potentials of the DNA-binding domains within the full-length HMGB1 protein were largely negative despite the presence of many positively charged residues. Our NMR data on counterions and electrostatic potentials show that the D/E repeats and DNA have similar electrostatic properties and compete for the DNA-binding domains. The competition promotes dissociation of the protein–DNA complex and influences the molecular behavior of the HMGB1 protein. These effects may be general among the DNA/RNA-binding proteins with D/E repeats.

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HMGB1 as an extracellular pro-inflammatory cytokine: Implications for drug-induced organic damage

Yuan,

Guo,

et al. 2024

Cell Biol Toxicol

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Drug-induced organic damage encompasses various intricate mechanisms, wherein HMGB1, a non-histone chromosome-binding protein, assumes a significant role as a pivotal hub gene. The regulatory functions of HMGB1 within the nucleus and extracellular milieu are interlinked. HMGB1 exerts a crucial regulatory influence on key biological processes including cell survival, inflammatory regulation, and immune response. HMGB1 can be released extracellularly from the cell during these processes, where it functions as a pro-inflammation cytokine. HMGB1 interacts with multiple cell membrane receptors, primarily Toll-like receptors (TLRs) and receptor for advanced glycation end products (RAGE), to stimulate immune cells and trigger inflammatory response. The excessive or uncontrolled HMGB1 release leads to heightened inflammatory responses and cellular demise, instigating inflammatory damage or exacerbating inflammation and cellular demise in different diseases. Therefore, a thorough review on the significance of HMGB1 in drug-induced organic damage is highly important for the advancement of pharmaceuticals, ensuring their effectiveness and safety in treating inflammation as well as immune-related diseases. In this review, we initially outline the characteristics and functions of HMGB1, emphasizing their relevance in disease pathology. Then, we comprehensively summarize the prospect of HMGB1 as a promising therapeutic target for treating drug-induced toxicity. Lastly, we discuss major challenges and propose potential avenues for advancing the development of HMGB1-based therapeutics. Graphical Abstract Graphical Headlights (1) A comprehensive overview of the intricate relationship between HMGB1 and drug-induced organ toxicity is presented, accompanied by the corresponding treatment strategies. (2) The present study addresses significant obstacles and suggests potential strategies for furthering the progress of HMGB1-based therapeutics. (3) The research prospects of HMGB1 are also summarized.

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DNA-mediated proteolysis by neutrophil elastase enhances binding activities of the HMGB1 protein

Cited by 8 publications

References 71 publications

HMGB1 cleavage by complement C1s and its potent anti-inflammatory product

HMGB1 cleavage by complement C1s and its potent anti-inflammatory product

Influence of an Intrinsically Disordered Region on Protein Domains Revealed by NMR-Based Electrostatic Potential Measurements

HMGB1 as an extracellular pro-inflammatory cytokine: Implications for drug-induced organic damage

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