Geiss-Friedlander R

References (13)

Title : N-terminal processing by dipeptidyl peptidase 9: Cut and Go! - Zolg_2024_Biochimie__
Author(s) : Zolg S , Donzelli L , Geiss-Friedlander R
Ref : Biochimie , : , 2024
Abstract : Dipeptidyl peptidase 9 (DPP9) is an intracellular amino-dipeptidase with physiological roles in the immune system, DNA repair and mitochondria homeostasis, while its deregulation is linked to cancer progression and immune-associated defects. Through its rare ability to cleave a peptide bond following the imino-acid proline, DPP9 acts as a molecular switch that regulates key proteins, such as the tumor-suppressor BRCA2. In this review we will discuss key concepts underlying the outcomes of protein processing by DPP9, including substrate turn-over by the N-degron pathway. Additionally, we will review non-enzymatic roles and the regulation of DPP9 by discussing the interactome of this protease, which includes SUMO1, Filamin A, NLRP1 and CARD8.
ESTHER : Zolg_2024_Biochimie__
PubMedSearch : Zolg_2024_Biochimie__
PubMedID: 38461970

Title : Chemoproteomics-Enabled Identification of 4-Oxo-beta-Lactams as Inhibitors of Dipeptidyl Peptidases 8 and 9 - Carvalho_2022_Angew.Chem.Int.Ed.Engl_61_e202210498
Author(s) : Carvalho LAR , Ross B , Fehr L , Bolgi O , Wohrle S , Lum KM , Podlesainski D , Vieira AC , Kiefersauer R , Felix R , Rodrigues T , Lucas SD , Gross O , Geiss-Friedlander R , Cravatt BF , Huber R , Kaiser M , Moreira R
Ref : Angew Chem Int Ed Engl , : , 2022
Abstract : Dipeptidyl peptidases 8 and 9 (DPP8/9) have gathered interest as drug targets due to their important roles in biological processes like immunity and tumorigenesis. Elucidation of their distinct individual functions remains an ongoing task and could benefit from the availability of novel, chemically diverse and selective chemical tools. Here, we report the activity-based protein profiling (ABPP)-mediated discovery of 4-oxo-beta-lactams as potent, non-substrate-like nanomolar DPP8/9 inhibitors. X-ray crystallographic structures revealed different ligand binding modes for DPP8 and DPP9, including an unprecedented targeting of an extended S2' (eS2') subsite in DPP8. Biological assays confirmed inhibition at both target and cellular levels. Altogether, our integrated chemical proteomics and structure-guided small molecule design approach led to novel DPP8/9 inhibitors with alternative molecular inhibition mechanisms, delivering the highest selectivity index reported to date.
ESTHER : Carvalho_2022_Angew.Chem.Int.Ed.Engl_61_e202210498
PubMedSearch : Carvalho_2022_Angew.Chem.Int.Ed.Engl_61_e202210498
PubMedID: 36089535
Gene_locus related to this paper: human-DPP8 , human-DPP9

Title : Dipeptidyl peptidase 9 triggers BRCA2 degradation and promotes DNA damage repair - Bolgi_2022_EMBO.Rep__e54136
Author(s) : Bolgi O , Silva-Garcia M , Ross B , Pilla E , Kari V , Killisch M , Spitzner M , Stark N , Lenz C , Weiss K , Donzelli L , Gorrell MD , Grade M , Riemer J , Urlaub H , Dobbelstein M , Huber R , Geiss-Friedlander R
Ref : EMBO Rep , :e54136 , 2022
Abstract : N-terminal sequences are important sites for post-translational modifications that alter protein localization, activity, and stability. Dipeptidyl peptidase 9 (DPP9) is a serine aminopeptidase with the rare ability to cleave off N-terminal dipeptides with imino acid proline in the second position. Here, we identify the tumor-suppressor BRCA2 as a DPP9 substrate and show this interaction to be induced by DNA damage. We present crystallographic structures documenting intracrystalline enzymatic activity of DPP9, with the N-terminal Met1-Pro2 of a BRCA21-40 peptide captured in its active site. Intriguingly, DPP9-depleted cells are hypersensitive to genotoxic agents and are impaired in the repair of DNA double-strand breaks by homologous recombination. Mechanistically, DPP9 targets BRCA2 for degradation and promotes the formation of RAD51 foci, the downstream function of BRCA2. N-terminal truncation mutants of BRCA2 that mimic a DPP9 product phenocopy reduced BRCA2 stability and rescue RAD51 foci formation in DPP9-deficient cells. Taken together, we present DPP9 as a regulator of BRCA2 stability and propose that by fine-tuning the cellular concentrations of BRCA2, DPP9 alters the BRCA2 interactome, providing a possible explanation for DPP9's role in cancer.
ESTHER : Bolgi_2022_EMBO.Rep__e54136
PubMedSearch : Bolgi_2022_EMBO.Rep__e54136
PubMedID: 35912982
Gene_locus related to this paper: human-DPP8 , human-DPP9

Title : Aerosol-based ligand soaking of reservoir-free protein crystals - Ross_2021_J.Appl.Crystallogr_54_895
Author(s) : Ross B , Krapp S , Geiss-Friedlander R , Littmann W , Huber R , Kiefersauer R
Ref : J Appl Crystallogr , 54 :895 , 2021
Abstract : Soaking of macromolecular crystals allows the formation of complexes via diffusion of molecules into a preformed crystal for structural analysis. Soaking offers various advantages over co-crystallization, e.g. small samples and high-throughput experimentation. However, this method has disadvantages, such as inducing mechanical stress on crystals and reduced success rate caused by low affinity/solubility of the ligand. To bypass these issues, the Picodropper was previously developed in the authors' laboratory. This technique aimed to deliver small volumes of compound solution in response to crystal dehydration supported by the Free Mounting System humidity control or by IR-laser-induced protein crystal transformation. Herein, a new related soaking development, the Aerosol-Generator, is introduced. This device delivers compounds onto the solution-free surface of protein crystals using an ultrasonic technique. The produced aerosol stream enables an easier and more accurate control of solution volumes, reduced crystal handling, and crystal-size-independent soaking. The Aerosol-Generator has been used to produce complexes of DPP8 crystals, where otherwise regular soaking did not achieve complex formation. These results demonstrate the potential of this device in challenging ligand-binding scenarios and contribute to further understanding of DPP8 inhibitor binding.
ESTHER : Ross_2021_J.Appl.Crystallogr_54_895
PubMedSearch : Ross_2021_J.Appl.Crystallogr_54_895
PubMedID: 34188616
Gene_locus related to this paper: human-DPP8

Title : Dipeptidyl peptidase 9 triggers BRCA2 degradation by the N-degron pathway to promote DNA-damage repair - Silva-Garcia_2020_Biorxiv__
Author(s) : Silva-Garcia M , Bolgi O , Ross B , Pilla E , Kari V , Killisch M , Stark N , Lenz C , Spitzner M , Gorrell MD , Grade M , Urlaub H , Dobbelstein M , Huber R , Geiss-Friedlander R
Ref : Biorxiv , : , 2020
Abstract : Dipeptidyl peptidase 9 (DPP9) is a serine protease cleaving N-terminal dipeptides preferentially post-proline with (patho)physiological roles in the immune system and cancer. Only few DPP9 substrates are known. Here we identify an association of human DPP9 with the tumour suppressor BRCA2, a key player in repair of DNA double-strand breaks that promotes the formation of RAD51 filaments. This interaction is triggered by DNA-damage and requires access to the DPP9 active-site. We present crystallographic structures documenting the N-terminal Met1-Pro2 of a BRCA21-40 peptide captured in the DPP9 active-site. Mechanistically, DPP9 targets BRCA2 for degradation by the N-degron pathway, and promotes RAD51 foci formation. Both processes are phenocopied by BRCA2 N-terminal truncation mutants, indicating that DPP9 regulates both stability and the cellular stoichiometric interactome of BRCA2. Consistently, DPP9-deprived cells are hypersensitive to DNA-damage. Together, we identify DPP9 as a regulator of BRCA2, providing a possible explanation for DPP9 involvement in cancer development.
ESTHER : Silva-Garcia_2020_Biorxiv__
PubMedSearch : Silva-Garcia_2020_Biorxiv__
PubMedID:
Gene_locus related to this paper: human-DPP8 , human-DPP9

Title : DPP8\/DPP9 inhibition elicits canonical Nlrp1b inflammasome hallmarks in murine macrophages - de Vasconcelos_2019_Life.Sci.Alliance_2_
Author(s) : de Vasconcelos NM , Vliegen G , Goncalves A , De Hert E , Martin-Perez R , Van Opdenbosch N , Jallapally A , Geiss-Friedlander R , Lambeir AM , Augustyns K , Van der Veken P , De Meester I , Lamkanfi M
Ref : Life Sciences Alliance , 2 : , 2019
Abstract : Activating germline mutations in the human inflammasome sensor NLRP1 causes palmoplantar dyskeratosis and susceptibility to Mendelian autoinflammatory diseases. Recent studies have shown that the cytosolic serine dipeptidyl peptidases DPP8 and DPP9 suppress inflammasome activation upstream of NLRP1 and CARD8 in human keratinocytes and peripheral blood mononuclear cells. Moreover, pharmacological inhibition of DPP8/DPP9 protease activity was shown to induce pyroptosis in murine C57BL/6 macrophages without eliciting other inflammasome hallmark responses. Here, we show that DPP8/DPP9 inhibition in macrophages that express a Bacillus anthracis lethal toxin (LeTx)-sensitive Nlrp1b allele triggered significantly accelerated pyroptosis concomitant with caspase-1 maturation, ASC speck assembly, and secretion of mature IL-1beta and IL-18. Genetic ablation of ASC prevented DPP8/DPP9 inhibition-induced caspase-1 maturation and partially hampered pyroptosis and inflammasome-dependent cytokine release, whereas deletion of caspase-1 or gasdermin D triggered apoptosis in the absence of IL-1beta and IL-18 secretion. In conclusion, blockade of DPP8/DPP9 protease activity triggers rapid pyroptosis and canonical inflammasome hallmarks in primary macrophages that express a LeTx-responsive Nlrp1b allele.
ESTHER : de Vasconcelos_2019_Life.Sci.Alliance_2_
PubMedSearch : de Vasconcelos_2019_Life.Sci.Alliance_2_
PubMedID: 30718379
Gene_locus related to this paper: human-DPP8 , human-DPP9

Title : Structures and mechanism of dipeptidyl peptidases 8 and 9, important players in cellular homeostasis and cancer - Ross_2018_Proc.Natl.Acad.Sci.U.S.A_115_E1437
Author(s) : Ross B , Krapp S , Augustin M , Kierfersauer R , Arciniega M , Geiss-Friedlander R , Huber R
Ref : Proc Natl Acad Sci U S A , 115 :E1437 , 2018
Abstract : Dipeptidyl peptidases 8 and 9 are intracellular N-terminal dipeptidyl peptidases (preferentially postproline) associated with pathophysiological roles in immune response and cancer biology. While the DPP family member DPP4 is extensively characterized in molecular terms as a validated therapeutic target of type II diabetes, experimental 3D structures and ligand-/substrate-binding modes of DPP8 and DPP9 have not been reported. In this study we describe crystal and molecular structures of human DPP8 (2.5 A) and DPP9 (3.0 A) unliganded and complexed with a noncanonical substrate and a small molecule inhibitor, respectively. Similar to DPP4, DPP8 and DPP9 molecules consist of one beta-propeller and alpha/beta hydrolase domain, forming a functional homodimer. However, they differ extensively in the ligand binding site structure. In intriguing contrast to DPP4, where liganded and unliganded forms are closely similar, ligand binding to DPP8/9 induces an extensive rearrangement at the active site through a disorder-order transition of a 26-residue loop segment, which partially folds into an alpha-helix (R-helix), including R160/133, a key residue for substrate binding. As vestiges of this helix are also seen in one of the copies of the unliganded form, conformational selection may contributes to ligand binding. Molecular dynamics simulations support increased flexibility of the R-helix in the unliganded state. Consistently, enzyme kinetics assays reveal a cooperative allosteric mechanism. DPP8 and DPP9 are closely similar and display few opportunities for targeted ligand design. However, extensive differences from DPP4 provide multiple cues for specific inhibitor design and development of the DPP family members as therapeutic targets or antitargets.
ESTHER : Ross_2018_Proc.Natl.Acad.Sci.U.S.A_115_E1437
PubMedSearch : Ross_2018_Proc.Natl.Acad.Sci.U.S.A_115_E1437
PubMedID: 29382749
Gene_locus related to this paper: human-DPP8 , human-DPP9

Title : DPP9 is a novel component of the N-end rule pathway targeting the tyrosine kinase Syk - Justa-Schuch_2016_Elife_5_
Author(s) : Justa-Schuch D , Silva-Garcia M , Pilla E , Engelke M , Kilisch M , Lenz C , Moller U , Nakamura F , Urlaub H , Geiss-Friedlander R
Ref : Elife , 5 : , 2016
Abstract : The aminopeptidase DPP9 removes dipeptides from N-termini of substrates having a proline or alanine in second position. Although linked to several pathways including cell survival and metabolism, the molecular mechanisms underlying these outcomes are poorly understood. We identified a novel interaction of DPP9 with Filamin A, which recruits DPP9 to Syk, a central kinase in B-cell signalling. Syk signalling can be terminated by degradation, requiring the ubiquitin E3 ligase Cbl. We show that DPP9 cleaves Syk to produce a neo N-terminus with serine in position 1. Pulse-chases combined with mutagenesis studies reveal that Ser1 strongly influences Syk stability. Furthermore, DPP9 silencing reduces Cbl interaction with Syk, suggesting that DPP9 processing is a prerequisite for Syk ubiquitination. Consistently, DPP9 inhibition stabilizes Syk, thereby modulating Syk signalling. Taken together, we demonstrate DPP9 as a negative regulator of Syk and conclude that DPP9 is a novel integral aminopeptidase of the N-end rule pathway.
ESTHER : Justa-Schuch_2016_Elife_5_
PubMedSearch : Justa-Schuch_2016_Elife_5_
PubMedID: 27614019

Title : Identification of novel dipeptidyl peptidase 9 substrates by two-dimensional differential in-gel electrophoresis - Zhang_2015_FEBS.J_282_3737
Author(s) : Zhang H , Maqsudi S , Rainczuk A , Duffield N , Lawrence J , Keane FM , Justa-Schuch D , Geiss-Friedlander R , Gorrell MD , Stephens AN
Ref : Febs J , 282 :3737 , 2015
Abstract : Dipeptidyl peptidase 9 (DPP9) is a member of the S9B/DPPIV (DPP4) serine protease family, which cleaves N-terminal dipeptides at an Xaa-Pro consensus motif. Cytoplasmic DPP9 has roles in epidermal growth factor signalling and in antigen processing, whilst the role of the recently discovered nuclear form of DPP9 is unknown. Mice lacking DPP9 proteolytic activity die as neonates. We applied a modified 2D differential in-gel electrophoresis approach to identify novel DPP9 substrates, using mouse embryonic fibroblasts lacking endogenous DPP9 activity. A total of 111 potential new DPP9 substrates were identified, with nine proteins/peptides confirmed as DPP9 substrates by MALDI-TOF or immunoblotting. Moreover, we also identified the dipeptide Val-Ala as a consensus site for DPP9 cleavage that was not recognized by DPP8, suggesting different in vivo roles for these closely related enzymes. The relative kinetics for the cleavage of these nine candidate substrates by DPP9, DPP8 and DPP4 were determined. This is the first identification of DPP9 substrates from cells lacking endogenous DPP9 activity. These data greatly expand the potential roles of DPP9 and suggest different in vivo roles for DPP9 and DPP8.
ESTHER : Zhang_2015_FEBS.J_282_3737
PubMedSearch : Zhang_2015_FEBS.J_282_3737
PubMedID: 26175140

Title : The amino terminus extension in the long dipeptidyl peptidase 9 isoform contains a nuclear localization signal targeting the active peptidase to the nucleus - Justa-Schuch_2014_Cell.Mol.Life.Sci_71_3611
Author(s) : Justa-Schuch D , Moller U , Geiss-Friedlander R
Ref : Cell Mol Life Sciences , 71 :3611 , 2014
Abstract : The intracellular prolyl peptidase DPP9 is implied to be involved in various cellular pathways including amino acid recycling, antigen maturation, cellular homeostasis, and viability. Interestingly, the major RNA transcript of DPP9 contains two possible translation initiation sites, which could potentially generate a longer (892 aa) and a shorter version (863 aa) of DPP9. Although the endogenous expression of the shorter DPP9 form has been previously verified, it is unknown whether the longer version is expressed, and what is its biological significance. By developing specific antibodies against the amino-terminal extension of the putative DPP9-long form, we demonstrate for the first time the endogenous expression of this longer isoform within cells. Furthermore, we show that DPP9-long represents a significant fraction of total DPP9 in cells, under steady-state conditions. Using biochemical cell fractionation assays in combination with immunofluorescence studies, we find the two isoforms localize to separate subcellular compartments. Whereas DPP9-short is present in the cytosol, DPP9-long localizes preferentially to the nucleus. This differential localization is attributed to a classical monopartite nuclear localization signal (K(K/R)X(K/R)) in the N-terminal extension of DPP9-long. Furthermore, we detect prolyl peptidase activity in nuclear fractions, which can be inhibited by specific DPP8/9 inhibitors. In conclusion, a considerable fraction of DPP9, which was previously considered as a purely cytosolic peptidase, localizes to the nucleus and is active there, raising the intriguing possibility that the longer DPP9 isoform may regulate the activity or stability of nuclear proteins, such as transcription factors.
ESTHER : Justa-Schuch_2014_Cell.Mol.Life.Sci_71_3611
PubMedSearch : Justa-Schuch_2014_Cell.Mol.Life.Sci_71_3611
PubMedID: 24562348

Title : The SUMO1-E67 Interacting Loop Peptide Is an Allosteric Inhibitor of the Dipeptidyl Peptidases 8 and 9 - Pilla_2013_J.Biol.Chem_288_32787
Author(s) : Pilla E , Kilisch M , Lenz C , Urlaub H , Geiss-Friedlander R
Ref : Journal of Biological Chemistry , 288 :32787 , 2013
Abstract : The intracellular peptidases dipeptidyl peptidase (DPP) 8 and DPP9 are involved in multiple cellular pathways including antigen maturation, cellular homeostasis, energy metabolism, and cell viability. Previously we showed that the small ubiquitin-like protein modifier SUMO1 interacts with an armlike structure in DPP9, leading to allosteric activation of the peptidase. Here we demonstrate that the E67-interacting loop (EIL) peptide, which corresponds to the interaction surface of SUMO1 with DPP9, acts as a noncompetitive inhibitor of DPP9. Moreover, by analyzing the sensitivity of DPP9 arm mutants to the EIL peptide, we mapped specific residues in the arm that are important for inhibition by the EIL, suggesting that the peptide acts as an allosteric inhibitor of DPP9. By modifying the EIL peptide, we constructed peptide variants with more than a 1,000-fold selectivity toward DPP8 (147 nm) and DPP9 (170 nm) over DPPIV (200 mum). Furthermore, application of these peptides to cells leads to a clear inhibition of cellular prolyl peptidase activity. Importantly, in line with previous publications, inhibition of DPP9 with these novel allosteric peptide inhibitors leads to an increase in EGF-mediated phosphorylation of Akt. This work highlights the potential use of peptides that mimic interaction surfaces for modulating enzyme activity.
ESTHER : Pilla_2013_J.Biol.Chem_288_32787
PubMedSearch : Pilla_2013_J.Biol.Chem_288_32787
PubMedID: 24072711
Gene_locus related to this paper: human-DPP8 , human-DPP9

Title : A Novel SUMO1-specific Interacting Motif in Dipeptidyl Peptidase 9 (DPP9) That Is Important for Enzymatic Regulation - Pilla_2012_J.Biol.Chem_287_44320
Author(s) : Pilla E , Moller U , Sauer G , Mattiroli F , Melchior F , Geiss-Friedlander R
Ref : Journal of Biological Chemistry , 287 :44320 , 2012
Abstract : Sumoylation affects many cellular processes by regulating the interactions of modified targets with downstream effectors. Here we identified the cytosolic dipeptidyl peptidase 9 (DPP9) as a SUMO1 interacting protein. Surprisingly, DPP9 binds to SUMO1 independent of the well known SUMO interacting motif, but instead interacts with a loop involving Glu(67) of SUMO1. Intriguingly, DPP9 selectively associates with SUMO1 and not SUMO2, due to a more positive charge in the SUMO1-loop. We mapped the SUMO-binding site of DPP9 to an extended arm structure, predicted to directly flank the substrate entry site. Importantly, whereas mutants in the SUMO1-binding arm are less active compared with wild-type DPP9, SUMO1 stimulates DPP9 activity. Consistent with this, silencing of SUMO1 leads to a reduced cytosolic prolyl-peptidase activity. Taken together, these results suggest that SUMO1, or more likely, a sumoylated protein, acts as an allosteric regulator of DPP9.
ESTHER : Pilla_2012_J.Biol.Chem_287_44320
PubMedSearch : Pilla_2012_J.Biol.Chem_287_44320
PubMedID: 23152501
Gene_locus related to this paper: human-DPP9

Title : The cytoplasmic peptidase DPP9 is rate-limiting for degradation of proline-containing peptides - Geiss-Friedlander_2009_J.Biol.Chem_284_27211
Author(s) : Geiss-Friedlander R , Parmentier N , Moller U , Urlaub H , Van den Eynde BJ , Melchior F
Ref : Journal of Biological Chemistry , 284 :27211 , 2009
Abstract : Protein degradation is an essential process that continuously takes place in all living cells. Regulated degradation of most cellular proteins is initiated by proteasomes, which produce peptides of varying length. These peptides are rapidly cleaved to single amino acids by cytoplasmic peptidases. Proline-containing peptides pose a specific problem due to structural constrains imposed by the pyrrolidine ring that prevents most peptidases from cleavage. Here we show that DPP9, a poorly characterized cytoplasmic prolyl-peptidase, is rate-limiting for destruction of proline-containing substrates both in cell extracts and in intact cells. We identified the first natural substrate for DPP9, the RU1(34-42) antigenic peptide (VPYGSFKHV). RU1(34-42) is degraded in vitro by DPP9, and down-regulation of DPP9 in intact cells results in increased presentation of this antigen. Together our findings demonstrate an important role for DPP9 in peptide turnover and antigen presentation.
ESTHER : Geiss-Friedlander_2009_J.Biol.Chem_284_27211
PubMedSearch : Geiss-Friedlander_2009_J.Biol.Chem_284_27211
PubMedID: 19667070
Gene_locus related to this paper: human-DPP9