DOCSLIB.ORG

Dipeptidase Aminodipeptidase, Quiescent Cell Proline of a Novel

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Dipeptidase Aminodipeptidase, Quiescent Cell Proline of a Novel Vesicular Localization and Characterization of a Novel Post-Proline-Cleaving Aminodipeptidase, Quiescent Cell Proline Dipeptidase This information is current as of September 26, 2021. Murali Chiravuri, Fernando Agarraberes, Suzanne L. Mathieu, Henry Lee and Brigitte T. Huber J Immunol 2000; 165:5695-5702; ; doi: 10.4049/jimmunol.165.10.5695 http://www.jimmunol.org/content/165/10/5695 Downloaded from References This article cites 28 articles, 18 of which you can access for free at: http://www.jimmunol.org/content/165/10/5695.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 26, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2000 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Vesicular Localization and Characterization of a Novel Post-Proline-Cleaving Aminodipeptidase, Quiescent Cell Proline Dipeptidase1 Murali Chiravuri,* Fernando Agarraberes,† Suzanne L. Mathieu,* Henry Lee,* and Brigitte T. Huber2* A large number of chemokines, cytokines, and signal peptides share a highly conserved X-Pro motif on the N-terminus. The cleavage of this N-terminal X-Pro dipeptide results in functional alterations of chemokines such as RANTES, stroma-derived factor-1, and macrophage-derived chemokine. Until recently, CD26/DPPIV was the only known protease with the ability to cleave N-terminal X-Pro motifs at neutral pH. We have isolated and cloned a novel serine protease, quiescent cell proline dipeptidase (QPP), with substrate specificity similar to that of CD26/DPPIV. In this paper we show that QPP, like CD26/DPPIV, is synthesized Downloaded from with a propeptide and undergoes N-glycosylation. Interestingly, this glycosylation is required for QPP enzymatic activity, but not for its localization. Unlike the cell surface molecule, CD26/DPPIV, QPP is targeted to intracellular vesicles that are distinct from lysosomes. Proteinase K treatment of intact vesicles indicates that QPP is located within the vesicles. These vesicles appear to have a secretory component, as QPP is secreted in a functionally active form in response to calcium release. The presence of QPP in the vesicular compartment suggests that molecules bearing the N-terminal X-Pro motif can be cleaved at multiple sites within and outside the cell. These results expand the potential site(s) and scope of a process that appears to be an important mechanism of http://www.jimmunol.org/ post-translational regulation. The Journal of Immunology, 2000, 165: 5695–5702. roteolytic cleavage of proteins can have profound effects C-terminal amino acid by prolylcarboxypeptidase (PCP; angio- on their stability and function (1–3). There is a striking tensinase C) (7, 8). P conservation of X-Pro motifs on the N termini of many Few proteolytic enzymes that cleave peptide bonds containing chemokines, cytokines, and other signal peptides, suggesting a se- proline have been identified. These include exopeptidases such as lective pressure for this motif in the evolution of these molecules CD26/DPPIV and PCP (4). We have isolated and cloned a serine (4). For many of these factors, the significance of the X-Pro motif protease, quiescent cell proline dipeptidase (QPP), which has sub- has yet to be ascertained. Recently, however, the work of several strate specificity very similar to that of CD26/DPPIV (9). Both by guest on September 26, 2021 groups has shown that the N-terminal X-Pro motif of the chemo- these enzymes cleave N-terminal dipeptides when the penultimate 3 kines RANTES, stroma-derived factor-1␣ (SDF1␣), SDF1␤, amino acid is a proline or, less preferably, an alanine residue (9– eotaxin, and macrophage-derived chemokine (MDC) is cleaved by 11). Recently, we observed that highly specific aminodipeptidase dipeptidyl peptidase IV (DPPIV/CD26), resulting in an alteration inhibitors cause cell death in quiescent lymphocytes, but not in of their biological function (2, 3, 5, 6). Thus, it appears that the activated lymphocytes (12). QPP is the likely target of these in- N-terminal X-Pro motif on signal molecules may serve as a site for hibitors, suggesting that this protease plays an important role in post-translational modification and regulation. The cleavage of resting cells (12). However, unlike CD26/DPPIV, which is found proline-containing peptide bonds on the C terminus can also affect on the cell surface (10, 11), QPP was isolated from intracellular the function of molecules. Angiotensin, for example, undergoes a fractions (9). Although QPP and CD26/DPPIV share substrate loss of vasoactive function following post-proline cleavage of the specificity, there is no significant sequence homology between these two proteases, suggesting an evolutionary convergence of *Department of Pathology, Program in Immunology, and †Department of Physiology, functional activity. QPP does, however, share significant sequence Sackler School of Graduate Biomedical Sciences, Tufts University School of Medi- cine, Boston, MA 02111 homology with the lysosomal serine protease PCP (42% sequence Received for publication March 15, 2000. Accepted for publication August 22, 2000. identity) (7, 9). It is interesting to note that the three post-proline- The costs of publication of this article were defrayed in part by the payment of page cleaving proteases, CD26/DPPIV, PCP, and QPP, all have the charges. This article must therefore be hereby marked advertisement in accordance same sequential ordering of the active site residues, Ser-Asp-His with 18 U.S.C. Section 1734 solely to indicate this fact. (4, 9), and are functionally active as homodimers (7, 9, 11). 1 This work was supported by National Institutes of Health Research Grants AI36696 In this work we show that similar to its functional homologue, and AI43469 (to B.T.H.). CD26/DPPIV, QPP is synthesized with a cleaved pro-peptide. 2 Address correspondence and reprint requests to Dr. Brigitte T. Huber, Department of Pathology, Tufts University School of Medicine, 136 Harrison Avenue, Boston, QPP undergoes N-linked glycosylation at the Golgi apparatus, MA 02111. E-mail address: [email protected] which is required for its proteolytic activity. Unlike CD26/DPPIV, 3 Abbreviations used in this paper: SDF, stroma-derived factor; QPP, quiescent cell QPP localizes to a post-Golgi vesicular compartment. QPP-con- proline dipeptidase; DPPIV, dipeptidyl peptidase IV; DPPII, dipeptidyl peptidase II; taining vesicles have similar density to lysosomes, but further frac- PCP, prolylcarboxypeptidase; GFP, green fluorescent protein; M6P, mannose-6-phos- phate; LAMP1, lysosome-associated membrane protein-1; MDC, macrophage-de- tionation shows that QPP-containing vesicles form a distinct com- rived chemokine, TGN, trans-Golgi network; zGGL-AMC, z-glycine-glycine- partment, and unlike mannose-6-phosphate (M6P)-dependent leucine-aminomethylcoumarin; Ala-Pro-AFC, alanine-proline-aminofluorocoumarin; HA, hemagglutinin; PNS, postnuclear supernatant; ER, endoplasmic reticulum; Glc- lysosomal targeting (13), QPP localizes to vesicles by a M6P- NAc, N-acetylglucosamine; PNGase F, peptide-N-glycosidase F. independent mechanism. QPP is localized in the vesicular lumen, Copyright © 2000 by The American Association of Immunologists 0022-1767/00/$02.00 5696 CHARACTERIZATION OF A CD26/DPPIV-LIKE PROTEASE as it is not susceptible to proteinase K digestion of intact vesicles. Following this the samples were brought up to 1% Nonidet P-40 and 0.05 Functionally active QPP can also be detected in cell culture su- M sodium phosphate. PNGase F (500 U) was added and incubated for the pernatant, and its secretion can be induced by cell activation and indicated times at 37°C. The reaction was terminated by boiling the sample in 2ϫ reducing loading buffer for 5 min, followed by SDS-PAGE and calcium mobilization, suggesting that there is a secretory compo- immunoblot analysis. The controls were treated in the same manner except nent to the QPP-containing vesicular compartment. The study of for the addition of PNGase F. post-translational modification of N-terminal X-Pro-containing signal molecules has been a rapidly expanding field in the last few Confocal microscopy years. The discovery of a novel post-proline-cleaving aminodipep- 293T human fibroblasts were transfected with QPP-GFP or QPP-Myc, tidase in a distinct compartment from CD26/DPPIV broadens the plated on coverslips, pretreated with fibronectin (100 ␮g/ml), and blocked scope of post-proline-cleavage as a regulatory mechanism. with 1% BSA in PBS. These cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X in PBS. The primary Ab was incubated for1hat4°C, followed by three washes and incubation of the secondary Materials and Methods Ab, where applicable, for1hat4°C. The samples were washed and vi- Materials sualized by confocal microscopy. Anti-Myc Ab was purchased from PharMingen (San Diego, CA), and anti- Biochemical organelle fractionation hemagglutinin (anti-HA) Ab was purchased from BabCo (Berkeley, CA). The anti-lysosome-associated membrane protein-1 (anti-LAMP1) mAb The isolation of subcellular organelles on discontinuous density gradients H4A3 was obtained from the Developmental Studies Hybridoma Bank was performed as previously described (14). In addition, a continuous gra- ϫ 8 (Iowa City, IA), and the anti-calnexin mAb was purchased from StressGen dient fractionation was performed. Briefly, 1 10 cells were lysed by (Victoria, Canada). Rab11 and adaptin-␣ Abs were purchased from Trans- cavitation or Dounce homogenization (Kontes, Vineland, NJ) in 0.25 M duction Laboratories (Lexington, KY).
Load more

Recommended publications
  • Catalytic Properties and Inhibition of Proline-Specific Dipeptidyl Peptidases II, IV and VII
    UC Berkeley UC Berkeley Previously Published Works Title Catalytic properties and inhibition of proline-specific dipeptidyl peptidases II, IV and VII Permalink https://escholarship.org/uc/item/4nf146zf Journal Biochemical Journal, 371 ISSN 0264-6021 Authors Leiting, B Pryor, K D Wu, J K et al. Publication Date 2003-04-01 Peer reviewed eScholarship.org Powered by the California Digital Library University of California Biochem. J. (2003) 371, 525–532 (Printed in Great Britain) 525 Catalytic properties and inhibition of proline-specific dipeptidyl peptidases II, IV and VII Barbara LEITING*1, KellyAnn D. PRYOR*, Joseph K. WU*, Frank MARSILIO*, Reshma A. PATEL*, Charles S. CRAIK†, Jonathan A. ELLMAN‡, Richard T. CUMMINGS* and Nancy A. THORNBERRY* *Department of Metabolic Disorders, Merck Research Laboratories, Mail code RY50G-236, P.O. Box 2000, Rahway, NJ 07065, U.S.A., †Department of Pharmaceutical Chemistry, University of California, 513 Parnassus Avenue, San Francisco, CA 94143-0446, U.S.A., and ‡Department of Chemistry, University of California, Berkeley, CA 94720, U.S.A. There is currently intense interest in the emerging group of strates and inhibitors for these enzymes, a complete biochemical proline-specific dipeptidases, and their roles in the regulation profile of these enzymes was obtained. The pH profiles, substrate of biological processes. Dipeptidyl peptidase IV (DPP-IV) is specificities as determined by positional scanning, Michaelis– involved in glucose metabolism by contributing to the regulation Menten constants and inhibition profiles for DPP-VII and DPP- of glucagon family peptides and has emerged as a potential target II were shown to be virtually identical, strongly supporting the for the treatment of metabolic diseases.
    [Show full text]
  • Single-Cell RNA Sequencing Demonstrates the Molecular and Cellular Reprogramming of Metastatic Lung Adenocarcinoma
    ARTICLE https://doi.org/10.1038/s41467-020-16164-1 OPEN Single-cell RNA sequencing demonstrates the molecular and cellular reprogramming of metastatic lung adenocarcinoma Nayoung Kim 1,2,3,13, Hong Kwan Kim4,13, Kyungjong Lee 5,13, Yourae Hong 1,6, Jong Ho Cho4, Jung Won Choi7, Jung-Il Lee7, Yeon-Lim Suh8,BoMiKu9, Hye Hyeon Eum 1,2,3, Soyean Choi 1, Yoon-La Choi6,10,11, Je-Gun Joung1, Woong-Yang Park 1,2,6, Hyun Ae Jung12, Jong-Mu Sun12, Se-Hoon Lee12, ✉ ✉ Jin Seok Ahn12, Keunchil Park12, Myung-Ju Ahn 12 & Hae-Ock Lee 1,2,3,6 1234567890():,; Advanced metastatic cancer poses utmost clinical challenges and may present molecular and cellular features distinct from an early-stage cancer. Herein, we present single-cell tran- scriptome profiling of metastatic lung adenocarcinoma, the most prevalent histological lung cancer type diagnosed at stage IV in over 40% of all cases. From 208,506 cells populating the normal tissues or early to metastatic stage cancer in 44 patients, we identify a cancer cell subtype deviating from the normal differentiation trajectory and dominating the metastatic stage. In all stages, the stromal and immune cell dynamics reveal ontological and functional changes that create a pro-tumoral and immunosuppressive microenvironment. Normal resident myeloid cell populations are gradually replaced with monocyte-derived macrophages and dendritic cells, along with T-cell exhaustion. This extensive single-cell analysis enhances our understanding of molecular and cellular dynamics in metastatic lung cancer and reveals potential diagnostic and therapeutic targets in cancer-microenvironment interactions. 1 Samsung Genome Institute, Samsung Medical Center, Seoul 06351, Korea.
    [Show full text]
  • Cell Communication and Signaling Biomed Central
    Cell Communication and Signaling BioMed Central Review Open Access Extravasation of leukocytes in comparison to tumor cells Carina Strell and Frank Entschladen* Address: Institute of Immunology, Witten/Herdecke University, Stockumer Str. 10, 58448 Witten, Germany Email: Carina Strell - [email protected]; Frank Entschladen* - [email protected] * Corresponding author Published: 4 December 2008 Received: 18 November 2008 Accepted: 4 December 2008 Cell Communication and Signaling 2008, 6:10 doi:10.1186/1478-811X-6-10 This article is available from: http://www.biosignaling.com/content/6/1/10 © 2008 Strell and Entschladen; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract The multi-step process of the emigration of cells from the blood stream through the vascular endothelium into the tissue has been termed extravasation. The extravasation of leukocytes is fairly well characterized down to the molecular level, and has been reviewed in several aspects. Comparatively little is known about the extravasation of tumor cells, which is part of the hematogenic metastasis formation. Although the steps of the process are basically the same in leukocytes and tumor cells, i.e. rolling, adhesion, transmigration (diapedesis), the molecules that are involved are different. A further important difference is that leukocyte interaction with the endothelium changes the endothelial integrity only temporarily, whereas tumor cell interaction leads to an irreversible damage of the endothelial architecture. Moreover, tumor cells utilize leukocytes for their extravasation as linkers to the endothelium.
    [Show full text]
  • Genetic Polymorphism of the Angiotensin-Converting Enzyme (ACE) in Asthmatic Patients
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector RESPIRATORY MEDICINE (1998) 92, 1305-1310 Original Articles Genetic polymorphism of the angiotensin-converting enzyme (ACE) in asthmatic patients H. TOMITA, S. SATO, R. MATSUDA, N. OGISU, T. MORI, T. NIIMI AND S. SHIMIZU 2nd Department of Intemal Medicine, Nagoya City University Medical School, Nagoya, Japan Angiotensin-converting enzyme (ACE) inactivates bradykinin, substance P and neurokinin A, which are believed to play important roles in the pathogenesis of asthma, especially in neurogenic inflammation. It has recently been shown that an insertion (1)ldeletion (D) polymorphism in the ACE gene accounts for variation in serum ACE levels. There are thus three genotypes (insertion homozygote, II; deletion homozygote, DD; heterozygotes, DI). The serum ACE level with the DD type is reported to be about double that of the II type and intermediate in the DI case. In the present study, we examined whether asthma is linked with this ACE gene polymorphism. Seventy-one patients with asthma (27 males and 44 females) and 142 sex- and age-matched healthy controls were determined for their genotype by the polymerase chain reaction (PCR) method. Twenty-five asthmatics demonstrated the II type (352%) 37 the DI type (52.1%), and nine the DD type (12.7%). There were no significant differences in the distributions of genotypes and serum ACE levels between healthy controls and patients. No significant differences were evident in serum IgE levels, age at onset, proportion of atopic type patients and severity of asthma among the three genotypes.
    [Show full text]
  • Lysosomal Membrane Glycoproteins Bind Cholesterol and Contribute to Lysosomal Cholesterol Export
    1 Lysosomal membrane glycoproteins bind cholesterol and 2 contribute to lysosomal cholesterol export 3 4 Jian Li and Suzanne R. Pfeffer* 5 Department of Biochemistry, Stanford University School of Medicine 6 Stanford, CA USA 94305-5307 7 8 *Correspondence to: [email protected]. 9 10 Abstract: LAMP1 and LAMP2 proteins are highly abundant, ubiquitous, mammalian proteins 11 that line the lysosome limiting membrane, and protect it from lysosomal hydrolase action. 12 LAMP2 deficiency causes Danon’s disease, an X-linked hypertrophic cardiomyopathy. LAMP2 13 is needed for chaperone-mediated autophagy, and its expression improves tissue function in 14 models of aging. We show here that LAMP1 and LAMP2 bind cholesterol in a manner that 15 buries the cholesterol 3β-hydroxyl group; they also bind tightly to NPC1 and NPC2 proteins that 16 export cholesterol from lysosomes. Quantitation of cellular LAMP2 and NPC1 protein levels 17 suggest that LAMP proteins represent a significant cholesterol binding site at the lysosome 18 limiting membrane, and may signal cholesterol availability. Functional rescue experiments show 19 that the ability of LAMP2 to facilitate cholesterol export from lysosomes relies on its ability to 20 bind cholesterol directly. 21 22 23 Introduction 24 Eukaryotic lysosomes are acidic, membrane-bound organelles that contain proteases, lipases and 25 nucleases and degrade cellular components to regenerate catabolic precursors for cellular use (1- 26 3). Lysosomes are crucial for the degradation of substrates from the cytoplasm, as well as 27 membrane bound compartments derived from the secretory, endocytic, autophagic and 28 phagocytic pathways. The limiting membrane of lysosomes is lined with so-called lysosomal 29 membrane glycoproteins (LAMPs) that are comprised of a short cytoplasmic domain, a single 30 transmembrane span, and a highly, N- and O-glycosylated lumenal domain (4-6).
    [Show full text]
  • B Number Gene Name Mrna Intensity Mrna
    sample) total list predicted B number Gene name assignment mRNA present mRNA intensity Gene description Protein detected - Membrane protein membrane sample detected (total list) Proteins detected - Functional category # of tryptic peptides # of tryptic peptides # of tryptic peptides detected (membrane b0002 thrA 13624 P 39 P 18 P(m) 2 aspartokinase I, homoserine dehydrogenase I Metabolism of small molecules b0003 thrB 6781 P 9 P 3 0 homoserine kinase Metabolism of small molecules b0004 thrC 15039 P 18 P 10 0 threonine synthase Metabolism of small molecules b0008 talB 20561 P 20 P 13 0 transaldolase B Metabolism of small molecules chaperone Hsp70; DNA biosynthesis; autoregulated heat shock b0014 dnaK 13283 P 32 P 23 0 proteins Cell processes b0015 dnaJ 4492 P 13 P 4 P(m) 1 chaperone with DnaK; heat shock protein Cell processes b0029 lytB 1331 P 16 P 2 0 control of stringent response; involved in penicillin tolerance Global functions b0032 carA 9312 P 14 P 8 0 carbamoyl-phosphate synthetase, glutamine (small) subunit Metabolism of small molecules b0033 carB 7656 P 48 P 17 0 carbamoyl-phosphate synthase large subunit Metabolism of small molecules b0048 folA 1588 P 7 P 1 0 dihydrofolate reductase type I; trimethoprim resistance Metabolism of small molecules peptidyl-prolyl cis-trans isomerase (PPIase), involved in maturation of b0053 surA 3825 P 19 P 4 P(m) 1 GenProt outer membrane proteins (1st module) Cell processes b0054 imp 2737 P 42 P 5 P(m) 5 GenProt organic solvent tolerance Cell processes b0071 leuD 4770 P 10 P 9 0 isopropylmalate
    [Show full text]
  • Table 6. Putative Genes Involved in the Utilization of Carbohydrates in G
    Table 6. Putative genes involved in the utilization of carbohydrates in G. thermodenitrificans NG80-2 genome Carbohydrates* Enzymes Gene ID Glycerol Glycerol Kinase GT1216 Glycerol-3-phosphate dehydrogenase, aerobic GT2089 NAD(P)H-dependent glycerol-3-phosphate dehydrogenase GT2153 Enolase GT3003 2,3-bisphosphoglycerate-independentphosphoglycerate mutase GT3004 Triosephosphate isomerase GT3005 3-phosphoglycerate kinase GT3006 Glyceraldehyde-3-phosphate dehydrogenase GT3007 Phosphoglycerate mutase GT1326 Pyruvate kinase GT2663 L-Arabinose L-arabinose isomerase GT1795 L-ribulokinase GT1796 L-ribulose 5-phosphate 4-epimerase GT1797 D-Ribose Ribokinase GT3174 Transketolase GT1187 Ribose 5-phosphate epimerase GT3316 D-Xylose Xylose kinase GT1756 Xylose isomerase GT1757 D-Galactose Galactokinase GT2086 Galactose-1-phosphate uridyltransferase GT2084 UDP-glucose 4-epimerase GT2085 Carbohydrates* Enzymes Gene ID D-Fructose 1-phosphofructokinase GT1727 Fructose-1,6-bisphosphate aldolase GT1805 Fructose-1,6-bisphosphate aldolase type II GT3331 Triosephosphate isomerase GT3005 D-Mannose Mannnose-6 phospate isomelase GT3398 6-phospho-1-fructokinase GT2664 D-Mannitol Mannitol-1-phosphate dehydrogenase GT1844 N-Acetylglucosamine N-acetylglucosamine-6-phosphate deacetylase GT2205 N-acetylglucosamine-6-phosphate isomerase GT2204 D-Maltose Alpha-1,4-glucosidase GT0528, GT1643 Sucrose Sucrose phosphorylase GT3215 D-Trehalose Alpha-glucosidase GT1643 Glucose kinase GT2381 Inositol Myo-inositol catabolism protein iolC;5-dehydro-2- GT1807 deoxygluconokinase
    [Show full text]
  • Sensitization to the Lysosomal Cell Death Pathway by Oncogene- Induced Down-Regulation of Lysosome-Associated Membrane Proteins 1 and 2
    Research Article Sensitization to the Lysosomal Cell Death Pathway by Oncogene- Induced Down-regulation of Lysosome-Associated Membrane Proteins 1 and 2 Nicole Fehrenbacher,1 Lone Bastholm,2 Thomas Kirkegaard-Sørensen,1 Bo Rafn,1 Trine Bøttzauw,1 Christina Nielsen,1 Ekkehard Weber,3 Senji Shirasawa,4 Tuula Kallunki,1 and Marja Ja¨a¨ttela¨1 1Apoptosis Department and Centre for Genotoxic Stress Response, Institute for Cancer Biology, Danish Cancer Society; 2Institute of Molecular Pathology, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark; 3Institute of Physiological Chemistry, Medical Faculty, Martin-Luther-University Halle-Wittenberg, Halle, Germany; and 4Department of Cell Biology, School of Medicine, Fukuoka University, Fukuoka, Japan Abstract molecules of the cell to breakdown products available for Expression and activity of lysosomal cysteine cathepsins metabolic reuse (3, 4). Cathepsin proteases are among the best- correlate with the metastatic capacity and aggressiveness of studied lysosomal hydrolases. They are maximally active at the tumors. Here, we show that transformation of murine acidic pH of lysosomes (pH 4–5). However, many of them can be Y527F embryonic fibroblasts with v-H-ras or c-src changes the active at the neutral pH outside lysosomes, albeit with a decreased distribution, density, and ultrastructure of the lysosomes, efficacy and/or altered specificity (5). For example, transformation and tumor environment enhance the expression of lysosomal decreases the levels of lysosome-associated membrane pro- teins (LAMP-1 and LAMP-2) in an extracellular signal- cysteine cathepsins and increase their secretion into the extracel- regulated kinase (ERK)- and cathepsin-dependent manner, lular space (6). Once outside the tumor cells, cathepsins stimulate and sensitizes the cells to lysosomal cell death pathways angiogenesis, tumor growth, and invasion in murine cancer induced by various anticancer drugs (i.e., cisplatin, etoposide, models, thereby enhancing cancer progression (7, 8).
    [Show full text]
  • Partial Characterisation of Dipeptidyl Peptidase 4 (DP 4
    Partial Characterisation of Dipeptidyl Peptidase 4 (DP 4) for the Treatment of Type 2 Diabetes and its Identification as a novel Pharmaceutical Target for Stress-related Indications Beiträge zur Charakterisierung der Dipeptidylpeptidase 4 (DP 4) für die Behandlung von Typ-2-Diabetes und deren Identifizierung als ein neuartiges pharmazeutisches Ziel für stressvermittelte Indikationen Der Naturwissenschaftlichen Fakultät der Friedrich-Alexander-Universität Erlangen-Nürnberg zur Erlangung des Doktorgrades Dr. rer. nat. vorgelegt von M. Sc. (Biochemie) Leona Wagner aus Rheinfelden (Baden) Als Dissertation genehmigt von der Naturwissenschaftlichen Fakultät der Friedrich-Alexander-Universität Erlangen-Nürnberg Tag der mündlichen Prüfung: 23.10.2012 Vorsitzender der Promotionskommission: Prof. Dr. Rainer Fink Erstberichterstatter: Prof. Dr. med. Stephan von Hörsten Zweitberichterstatter: Prof. Dr. Johann Helmut Brandstätter Publications, Presentations and Posters Part of the present study have been published or submitted in the following journals or presented on the following conferences: Publications: 1. Hinke, S.A., McIntosh, C.H., Hoffmann, T., Kuhn-Wache, K., Wagner, L., Bar, J., Manhart, S., Wermann, M., Pederson, R.A., and Demuth, H.U. (2002) On combination therapy of diabetes with metformin and dipeptidyl peptidase IV inhibitors. Diabetes Care 25, 1490- 1491. 2. Bär, J., Weber, A., Hoffmann, T., Stork, J., Wermann, M., Wagner, L., Aust, S., Gerhartz, B., and Demuth, H.U. (2003) Characterisation of human dipeptidyl peptidase IV expressed in Pichia pastoris. A structural and mechanistic comparison between the recombinant human and the purified porcine enzyme. Biol. Chem. 384, 1553-1563. 3. Engel, M., Hoffmann, T., Wagner, L., Wermann, M., Heiser, U., Kiefersauer, R., Huber, R., Bode, W., Demuth, H.U., and Brandstetter, H.
    [Show full text]
  • Aminopeptidase P1 Recombinant Protein Cat
    Aminopeptidase P1 Recombinant Protein Cat. No.: 91-173 Aminopeptidase P1 Recombinant Protein Specifications SPECIES: Human SOURCE SPECIES: E. coli SEQUENCE: Pro2-His623 FUSION TAG: C-6 His tag TESTED APPLICATIONS: APPLICATIONS: This recombinant protein can be used for biological assays. For research use only. PREDICTED MOLECULAR 70.6 kD WEIGHT: Properties Greater than 95% as determined by reducing SDS-PAGE. PURITY: Endotoxin level less than 0.1 ng/ug (1 IEU/ug) as determined by LAL test. PHYSICAL STATE: Liquid Supplied as a 0.2 um filtered solution of 20mM TrisHCl, 10% Glycerol, pH 8.0. It is not BUFFER: recommended to reconstitute to a concentration less than 100 ug/ml. September 23, 2021 1 https://www.prosci-inc.com/aminopeptidase-p1-recombinant-protein-91-173.html Store at -20˚C, stable for 6 months after receipt. STORAGE CONDITIONS: Please aliquot the reconstituted solution to minimize freeze-thaw cycles. Additional Info OFFICIAL SYMBOL: XPNPEP1 Xaa-Pro Aminopeptidase 1, Aminoacylproline Aminopeptidase, Cytosolic Aminopeptidase ALTERNATE NAMES: P, Soluble Aminopeptidase P, sAmp, X-Pro Aminopeptidase 1, X-Prolyl Aminopeptidase 1 Soluble, XPNPEP1, XPNPEPL, XPNPEPL1 ACCESSION NO.: Q9NQW7 GENE ID: 7511 Background and References X-Prolyl Aminopeptidase (XPNPEP1) is a proline-specific metalloaminopeptidase that specifically catalyzes the removal of any unsubstituted N-terminal amino acid that is adjacent to a penultimate proline residue. Because of its specificity toward proline, it has been suggested that X-Prolyl Aminopeptidase is important in the maturation and degradation of peptide hormones, neuropeptides, and tachykinins, as well as in the digestion of otherwise resistant dietary protein fragments, thereby complementing the pancreatic peptidases.
    [Show full text]
  • Fluid Biomarkers in Frontotemporal Dementia: Past, Present and Future
    Neurodegeneration J Neurol Neurosurg Psychiatry: first published as 10.1136/jnnp-2020-323520 on 13 November 2020. Downloaded from Review Fluid biomarkers in frontotemporal dementia: past, present and future Imogen Joanna Swift ,1 Aitana Sogorb- Esteve,1,2 Carolin Heller ,1 Matthis Synofzik,3,4 Markus Otto ,5 Caroline Graff,6,7 Daniela Galimberti ,8,9 Emily Todd,2 Amanda J Heslegrave,1 Emma Louise van der Ende ,10 John Cornelis Van Swieten ,10 Henrik Zetterberg,1,11 Jonathan Daniel Rohrer 2 ► Additional material is ABSTRACT into a behavioural form (behavioural variant fron- published online only. To view, The frontotemporal dementia (FTD) spectrum of totemporal dementia (bvFTD)), a language variant please visit the journal online (primary progressive aphasia (PPA)) and a motor (http:// dx. doi. org/ 10. 1136/ neurodegenerative disorders includes a heterogeneous jnnp- 2020- 323520). group of conditions. However, following on from a series presentation (either FTD with amyotrophic lateral of important molecular studies in the early 2000s, major sclerosis (FTD- ALS) or an atypical parkinsonian For numbered affiliations see advances have now been made in the understanding disorder). Neuroanatomically, the FTD spectrum end of article. of the pathological and genetic underpinnings of the is characteristically associated with dysfunction and disease. In turn, alongside the development of novel neuronal loss in the frontal and temporal lobes, but Correspondence to more widespread cortical, subcortical, cerebellar Dr Jonathan Daniel Rohrer, methodologies for measuring proteins and other Dementia Research molecules in biological fluids, the last 10 years have and brainstem involvement is now recognised. Centre, Department of seen a huge increase in biomarker studies within FTD.
    [Show full text]
  • Brush Border and Cytosol Peptidase Activities of Human Small Intestine in Normal Subjects and Celiac Patients
    Pediatr. Res. 14: 8 12-8 18 ( 1980) brush border membrane digestive peptidase: celiac disease small intestine cytosol membrane Brush Border and Cytosol Peptidase Activities of Human Small Intestine in Normal Subjects and Celiac Patients CiLNtKOSO ANIIKIA. SALVIZ'TOKE <'UC'<'lIIAKA. BIZSILIO DL: VIZIA. (;IOKGIO IIL: KITIS. <;ABRIEL.E MAZZA<'<'A. AN11 SALVATORE 1Z~R1~('1110'~" I)c.prrrrn~enrof Pediclrrrc.~ond Depurrmenr of (;ii.crrt~c~nrerologr,I1 I.irc~ttlt~ij .&ledicrne. L'tt~r.i~rvrtrof Ni~pler.Sc~plt,\. Irull.. und ('on.\r,qlro .Vu:ronule dcdle Rrc.erchr. Pro~rirrn(4- Prevc,ttrrve Medic~itrc(Prt~~cc./ I'ermi~rirl ,Wedrc.itlc~. Rontc. Irul, Summary Studies in animals have demonstrated two major subcellular localization of digestive peptidases in the enterocyte. cytosol. and Peptidase activities have been investigated in the brush border brush border, M~~~ of the hydrolyzing dipep- of human proximal jejunum by using dipeptides and tripeptides [ides tripeprides is localized in ,he cytosol (1, 16, 17. 21, 26. and P-naphthylamides of glycyl-I.-proline and amino acids as sub- 32, 35, 42, 46, 49, 53, 54, 60) few except,ons (25, 26. 42. 46. \trates. I'he activities hydrolyzing glycyl-I.-leucine. I.-phenylalanyl- 49, 60): three soluble enzymes dipeptidase and tripeptidase I.-alaninc, and I.-le~cyl~lycylglycinein the brush horder were found have been and (12, 15, 16, 20, 48. to be only 1.5. 15. and 16% of the total peptidase activities present 5 1. 62). Almost all the hydrolyzing the P- in the intestinal mucosa, but the specific activities for the hydrol- naphthylamides ofamino (6.
    [Show full text]