Structural basis for the recognition and cleavage of histone H3 by cathepsin L

Adams-Cioaba, Melanie A.; Krupa, Joanne C.; Xu, Chao; Mort, John S.; Min, Jinrong

doi:10.1038/ncomms1204

Cited by 65 publications

(60 citation statements)

References 36 publications

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“…However, we cannot exclude that other proteases may also be involved in histone H3 proteolysis, 52 because intracellular proteases, including cathepsin L, are known to cleave H3 to a similarly sized band. [53][54][55] In addition, histone H3 is cleaved by glutamate dehydrogenase at the same position, also cleaving after K27. 56,57 Nonetheless, we did not observe an effect of H3 truncation on the cytotoxicity of nucleosomes.…”

Section: Discussionmentioning

confidence: 99%

DNA and factor VII–activating protease protect against the cytotoxicity of histones

et al. 2017

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Key Points• Free histones, not nucleosomes, are cytotoxic and are degraded by FSAP in serum to protect against cytotoxicity.• Free histone H3 was not detectable in sera of septic baboons and patients with meningococcal sepsis.Circulating histones have been implicated as major mediators of inflammatory disease because of their strong cytotoxic effects. Histones form the protein core of nucleosomes;however, it is unclear whether histones and nucleosomes are equally cytotoxic. Several plasma proteins that neutralize histones are present in plasma. Importantly, factor VII-activating protease (FSAP) is activated upon contact with histones and subsequently proteolyzes histones. We aimed to determine the effect of FSAP on the cytotoxicity of both histones and nucleosomes. Indeed, FSAP protected against histone-induced cytotoxicity of cultured cells in vitro. Upon incubation of serum with histones, endogenous FSAP was activated and degraded histones, which also prevented cytotoxicity. Notably, histones as part of nucleosome complexes were not cytotoxic, whereas DNA digestion restored cytotoxicity. Histones in nucleosomes were inefficiently cleaved by FSAP, which resulted in limited cleavage of histone H3 and removal of the N-terminal tail. The specific isolation of either circulating nucleosomes or free histones from sera of Escherichia coli challenged baboons or patients with meningococcal sepsis revealed that histone H3 was present in the form of nucleosomes, whereas free histone H3 was not detected. All samples showed signs of FSAP activation. Markedly, we observed that all histone H3 in nucleosomes from the patients with sepsis, and most histone H3 from the baboons, was N-terminally truncated, giving rise to a similarly sized protein fragment as through cleavage by FSAP.Taken together, our results suggest that DNA and FSAP jointly limit histone cytotoxicity and that free histone H3 does not circulate in appreciable concentrations in sepsis.

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Section: Discussionmentioning

confidence: 99%

DNA and factor VII–activating protease protect against the cytotoxicity of histones

et al. 2017

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“…Recently, the groups of Min and Mort made attempts to gain insight into the interactions between Abbreviations C25S, point mutation at 25th cysteine to serine using mature cathepsin L numbering; NAG, N-acetyl-D-glucosamine; SDS/PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis. a part of the physiological substrate histone H3 and human cathepsin L by crystallizing the complex with a catalytic mutant of the enzyme and elucidating its structure [16]. Refinement confirmed the positioning of only three amino acids from the histone H3 [19][20][21][22][23][24][25][26][27][28][29][30][31][32][33] peptide.…”

Section: Abstract: Cathepsin; Cysteine Cathepsin; Substrate Interactionmentioning

confidence: 99%

Caught in the act: the crystal structure of cleaved cathepsin L bound to the active site of Cathepsin L

Sosnowski

Turk

2016

FEBS Letters

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Cathepsin L is a ubiquitously expressed papain-like cysteine protease involved in the endosomal degradation of proteins and has numerous roles in physiological and pathological processes, such as arthritis, osteoporosis, and cancer. Insight into the specificity of cathepsin L is important for elucidating its physiological roles and drug discovery. To study interactions with synthetic ligands, we prepared a presumably inactive mutant and crystallized it. Unexpectedly, the crystal structure determined at 1.4 A revealed that the cathepsin L molecule is cleaved, with the cleaved region trapped in the active site cleft of the neighboring molecule. Hence, the catalytic mutant demonstrated low levels of catalytic activity.

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“…Identification of Histone H2A Clipping Enzyme by in Vitro proteolytic Assay-Given the large variety of truncated histone proteoforms identified, we further investigated whether cathepsin L, a protease responsible for histone H3 and H3.3 clipping (9,37,38), is capable of cleaving histone H2A. To do so, we performed an in vitro assay incubating recombinant histone H2A with cathepsin L and characterized the cleavage specificity using nano-LC coupled to an Orbitrap Fusion with FETD.…”

Section: Table I Truncated H2b Proteoforms Observed In Iipt/parallel mentioning

confidence: 99%

Analyses of Histone Proteoforms Using Front-end Electron Transfer Dissociation-enabled Orbitrap Instruments

Anderson

Karch

Ugrin

et al. 2016

Molecular & Cellular Proteomics

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Chromatin is the structural framework that packages DNA into chromosomes within the nucleus of a cell (2). Histones comprise the principal protein component of chromatin and are involved in the regulation of gene expression (3,4). This epigenetic regulation is achieved through complex patterns of post-translational modifications (PTMs), 1 the incorporation of histone variants, and through controlled histone proteolysis (5-10). Comprehensive characterization of histones by mass spectrometry (MS) has proven technically difficult for a number of reasons. Traditional methods (bottom-up MS) of sequence determination and PTM site localization are not practical. Histone N-terminal regions are rich in lysine and arginine residues, and thus proteolysis using trypsin generates peptides that are too small or that are poorly retained on reversephase HPLC C18 resins for subsequent MS detection (11). With the advent of electron transfer dissociation (ETD) and more efficient electron capture dissociation fragmentation methods, which are better suited for larger, more highly charged peptides (12, 13), several studies utilizing other endoproteases to generate longer peptides have emerged (14 -16). Although these methodologies do well to preserve the combinatorial PTM profiles of histone tails, in some cases it is still impossible to identify the proteoforms from which these peptides originate. This is why analyzing histones intact, as they exist in the cells from which they are derived, is the best method for identifying unique histone proteoforms.The results of several recent studies involving top-down analyses of histones highlight the complexity of the histone From the ‡Department

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Structural basis for the recognition and cleavage of histone H3 by cathepsin L

Cited by 65 publications

References 36 publications

DNA and factor VII–activating protease protect against the cytotoxicity of histones

DNA and factor VII–activating protease protect against the cytotoxicity of histones

Caught in the act: the crystal structure of cleaved cathepsin L bound to the active site of Cathepsin L

Analyses of Histone Proteoforms Using Front-end Electron Transfer Dissociation-enabled Orbitrap Instruments

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