DOCSLIB.ORG

Rapid Affinity Purification of Intracellular Organelles Using a Twin Strep Tag Jian Xiong1,2,*, Jingquan He1,*, Wendy P

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Rapid Affinity Purification of Intracellular Organelles Using a Twin Strep Tag Jian Xiong1,2,*, Jingquan He1,*, Wendy P © 2019. Published by The Company of Biologists Ltd | Journal of Cell Science (2019) 132, jcs235390. doi:10.1242/jcs.235390 TOOLS AND RESOURCES Rapid affinity purification of intracellular organelles using a twin strep tag Jian Xiong1,2,*, Jingquan He1,*, Wendy P. Xie1, Ezekiel Hinojosa1, Chandra Shekar R. Ambati3, Nagireddy Putluri3,4, Hyun-Eui Kim1,2, Michael X. Zhu1,2,‡ and Guangwei Du1,2,‡ ABSTRACT complex 1 (mTORC1) is recruited to and activated on the lysosomal Cells are internally organized into compartmentalized organelles that surface by sensing the abundance of nutrients in the lumen, such as execute specialized functions. To understand the functions of amino acids and cholesterol (Castellano et al., 2017; Zoncu et al., individual organelles and their regulations, it is critical to resolve the 2011). Similarly, mitochondria can also function as a signaling compositions of individual organelles, which relies on a rapid and organelle (Chandel, 2014). For example, cytochrome c released from efficient isolation method for specific organellar populations. Here, we the mitochondria initiates cell death (Bhola and Letai, 2016; Burke, introduce a robust affinity purification method for rapid isolation of 2017; Liu et al., 1996). Another example is AKAP family proteins, intracellular organelles (e.g. lysosomes, mitochondria and which anchor and regulate the activities of protein kinase A and other peroxisomes) by taking advantage of the extraordinarily high affinity signaling enzymes on the outer membrane of mitochondria (Chandel, between the twin strep tag and streptavidin variants. With this 2014; Esseltine and Scott, 2013). method, we can isolate desired organelles with high purity and yield in With rapid technical advancements, profiling the global levels of 3 min from the post-nuclear supernatant of mammalian cells or less RNA, protein, lipids and metabolites has become common in than 8 min for the whole purification process. Using lysosomes as an current biomedical research. However, most of these large-scale example, we show that the rapid procedure is especially useful for profiling studies do not provide spatial information (Uhlen et al., studying transient and fast cellular activities, such as organelle- 2015), thus cannot explain how different organelles regulate their initiated signaling and organellar contents of small-molecular highly compartmentalized cellular functions. The ability of metabolites. Therefore, our method offers a powerful tool to dissect measuring the compositions of specific organellar populations spatiotemporal regulation and functions of intracellular organelles. and their changes in response to stimuli would provide a powerful tool to understand the functions of these organelles. KEY WORDS: Twin strep tag, Lysosomes, Mitochondria, Isolation of different organelles is traditionally accomplished by Peroxisomes subcellular fractionation with differential centrifugation or multi- step density gradient ultracentrifugation (de Araujo and Huber, INTRODUCTION 2007; Foster et al., 2006; Frezza et al., 2007; Graham, 2001a,b,c; Eukaryotic cells are compartmentalized into distinct membrane- Michelsen and von Hagen, 2009). However, most subcellular enclosed organelles. Each organelle carries hundreds to thousands of fractionation approaches bear some intrinsic drawbacks. For proteins, lipids and metabolites, and performs a unique set of cellular example, the heterogeneous nature in the density of any given functions. For example, lysosomes are the major degradation organellar population makes it difficult to obtain a type of compartments responsible for the clearance of unnecessary organelle without contamination from the others. In addition, the materials inside the cells (Kurz et al., 2008). Mitochondria are the concentration of a desired organellar population collected from major organelles for the ATP generation required to support cellular multiple fractions is often relatively low, making some downstream synthetic pathways (Chandel, 2014). Peroxisomes are the primary analyses challenging. Moreover, to the best of our knowledge, the vesicles that catabolize long-chain fatty acids and regulate the balance subcellular fractionation methods usually take more than an hour of oxidization (Lodhi and Semenkovich, 2014; Smith and Aitchison, (Frezza et al., 2007; Graham, 2001a,b,c), which may lead to 2013). In addition to their well-known classic functions, recent changes in the compositions of organelles, especially the signaling studies have also revealed that some organelles are directly involved molecules associated with the cytoplasmic leaflet of the organelles in cell signaling. For example, mechanistic target of rapamycin and some labile small-molecule metabolites. Besides fractionation, specific methods have also been developed for the purification of certain organelles. For example, lysosomes can be isolated by 1Department of Integrative Biology and Pharmacology, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, TX 77030, magnets after being loaded with iron oxide-conjugated dextrans USA. 2Biochemistry and Cell Biology Program, MD Anderson Cancer Center (Rofe and Pryor, 2016). However, depending on the duration of UTHealth Graduate School of Biomedical Sciences, Houston, TX 77030, USA. 3Dan loading and chasing, dextrans are enriched to different degrees in L. Duncan Cancer Center, Advanced Technology Core, Alkek Center for Molecular Discovery, Baylor College of Medicine, Houston, TX 77030, USA. 4Department of various endosome populations and lysosomes (Humphries et al., Molecular & Cell Biology, Baylor College of Medicine, Houston, TX 77030, USA. 2011). Moreover, long-term accumulation of non-degradable *These authors contributed equally to this work dextran may have some unexpected effects on lysosomal ‡Authors for correspondence ([email protected]; guangwei.du@uth. functions (Kurz et al., 2008). Some recent studies have shown the tmc.edu) successful purification of mitochondria and lysosomes by using beads conjugated to antibodies against an endogenous J.H., 0000-0002-9341-8750; W.P.X., 0000-0002-0085-4415; M.X.Z., 0000-0002- 5676-841X; G.D., 0000-0003-4193-6975 mitochondrial or lysosomal-resident protein (Franko et al., 2013; Michelsen and von Hagen, 2009), or against an epitope tag fused to Received 17 June 2019; Accepted 15 November 2019 these resident proteins (Abu-Remaileh et al., 2017; Ahier et al., Journal of Cell Science 1 TOOLS AND RESOURCES Journal of Cell Science (2019) 132, jcs235390. doi:10.1242/jcs.235390 2018; Chen et al., 2016; Zoncu et al., 2011). Antibody affinity Furthermore, using lysosomes as an example, we demonstrate purification is fast and specific; thus it overcomes some drawbacks that this method can be used to monitor the transient lysosomal of the traditional approaches. However, antibody affinity association of signaling protein complexes as well as small- purification requires a substantial amount of antibody. In addition, molecule metabolites. the elution of functional organelles is achieved via the competition by a high concentration of epitope peptides, which is usually not RESULTS very efficient. Design of a new affinity lysosome purification approach One popular protein purification strategy is fusing the proteins of using twin strep tag interest to the Strep II tag (WSHPQFEK), which mimics the strong Many applications involved in organelle purification, such as interaction between biotin and streptavidin (Kimple et al., 2013; evaluation of signaling events and measurement of small-molecule Schmidt and Skerra, 2007). Strep II-tagged proteins can be metabolites, require rapid recovery as well as maintenance of efficiently eluted by a low concentration of biotin derivatives. The organelles in the physiological conditions during purification and small Strep II tag is biologically inert, and the binding between the elution. To test the use of twin strep tag in organelle purification, we tagged proteins and streptavidin beads can take place under fused the twin strep tag to the C-terminus of monomeric GFP physiological conditions. In addition to protein purification, the (mGFP)-fused LAMP1, a lysosome-resident protein (hereafter readily reversible interaction has allowed the use of Strep II tag for denoted Lyso-2Strep) (Fig. 1A). The cytoplasmic orientation of the purification of live antigen-specific T cells (Liu et al., 2016). the twin strep would allow subsequent purification of lysosomes The recently generated streptavidin variant, Strep-Tactin XT, using streptavidin beads (Fig. 1B). All lysosomal membrane further increases the binding affinity between two tandem Strep proteins are synthesized in the rough endoplasmic reticulum and tags (herein denoted the ‘twin strep tag’) (Schmidt et al., 2013; transported to trans-Golgi network before they are delivered to Yeliseev et al., 2017). As compared to micromolar-to-nanomolar lysosomes (Braulke and Bonifacino, 2009). The expression of Lyso- dissociation constant (Kd) for most commercial epitope antibodies, 2Strep is under the control of a tetracycline-inducible promoter, such as FLAG, Myc and HA (Einhauer and Jungbauer, 2001; which offers the option of turning off the transcription a few hours LaCava et al., 2015; Schiweck et al., 1997; Wegner et al., 2002), before lysosome isolation. This allows lysosomal delivery of newly twin strep tag features a nanomolar-to-picomolar Kd towards Strep- translated Lyso-2Strep protein, ensuring that the majority of Tactin XT
Load more

Recommended publications
  • Multiple Pathways for Protein Transport to Peroxisomes
    Review Multiple Pathways for Protein Transport to Peroxisomes P.K. Kim 1,2 and E.H. Hettema 3 1 - Program in Cell Biology, Hospital for Sick Children, Toronto, ON, Canada M5G 1X8 2 - Department of Biochemistry, University of Toronto, Toronto, ON, Canada M5S 1A8 3 - Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court, Western Bank, Sheffield, South Yorkshire S10 2TN, United Kingdom Correspondence to E.H. Hettema: [email protected] http://dx.doi.org/10.1016/j.jmb.2015.02.005 Edited by S. High Abstract Peroxisomes are unique among the organelles of the endomembrane system. Unlike other organelles that derive most if not all of their proteins from the ER (endoplasmic reticulum), peroxisomes contain dedicated machineries for import of matrix proteins and insertion of membrane proteins. However, peroxisomes are also able to import a subset of their membrane proteins from the ER. One aspect of peroxisome biology that has remained ill defined is the role the various import pathways play in peroxisome maintenance. In this review, we discuss the available data on matrix and membrane protein import into peroxisomes. © 2015 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). Introduction dependent on consumption of energy in the form of ATP. Several aspects of peroxisomal protein transport Peroxisomes are organelles found in almost all distinguish it from other translocation systems. For eukaryotic cells. They are bounded by a single example, peroxisomal proteins can fold, acquire membrane and are usually spherical.
    [Show full text]
  • Characterization of the Macrophage Transcriptome in Glomerulonephritis-Susceptible and -Resistant Rat Strains
    Genes and Immunity (2011) 12, 78–89 & 2011 Macmillan Publishers Limited All rights reserved 1466-4879/11 www.nature.com/gene ORIGINAL ARTICLE Characterization of the macrophage transcriptome in glomerulonephritis-susceptible and -resistant rat strains K Maratou1, J Behmoaras2, C Fewings1, P Srivastava1, Z D’Souza1, J Smith3, L Game4, T Cook2 and T Aitman1 1Physiological Genomics and Medicine Group, MRC Clinical Sciences Centre, Imperial College London, London, UK; 2Centre for Complement and Inflammation Research, Imperial College London, London, UK; 3Renal Section, Imperial College London, London, UK and 4Genomics Laboratory, MRC Clinical Sciences Centre, London, UK Crescentic glomerulonephritis (CRGN) is a major cause of rapidly progressive renal failure for which the underlying genetic basis is unknown. Wistar–Kyoto (WKY) rats show marked susceptibility to CRGN, whereas Lewis rats are resistant. Glomerular injury and crescent formation are macrophage dependent and mainly explained by seven quantitative trait loci (Crgn1–7). Here, we used microarray analysis in basal and lipopolysaccharide (LPS)-stimulated macrophages to identify genes that reside on pathways predisposing WKY rats to CRGN. We detected 97 novel positional candidates for the uncharacterized Crgn3–7. We identified 10 additional secondary effector genes with profound differences in expression between the two strains (45-fold change, o1% false discovery rate) for basal and LPS-stimulated macrophages. Moreover, we identified eight genes with differentially expressed alternatively spliced isoforms, by using an in-depth analysis at the probe level that allowed us to discard false positives owing to polymorphisms between the two rat strains. Pathway analysis identified several common linked pathways, enriched for differentially expressed genes, which affect macrophage activation.
    [Show full text]
  • Peroxin3, a Newly Identified Regulator of Melanocyte Development and Melanosome Biogenesis in Zebrafish Danio Rerio
    Peroxin3, a newly identified regulator of melanocyte development and melanosome biogenesis in zebrafish Danio rerio Dissertation zur Erlangung des Doktorgrades (Dr. rer. nat.) der Mathematisch-Naturwissenschaftlichen Fakultät der Rheinischen Friedrich-Wilhelms-Universität Bonn vorgelegt von Mirco Brondolin aus San Dona’ di Piave - Italien Bonn 2016 Angefertigt mit Genehmigung der Mathematisch-Naturwissenschaftlichen Fakultät der Rheinischen Friedrich-Wilhelms-Universität Bonn 1. Gutachter Prof. Dr. rer. nat. Michael Hoch 2. Gutachter Prof. Dr. phil. nat. Christoph Thiele Tag der Promotion: 20. März 2017 Erscheinungsjahr: 2017 Per aspera sic itur ad astra “Through hardships to the stars” (Lucius Annaeus Seneca, Hercules furens, act II, v. 437 Table of contents I Table of contents 1 Introduction ...........................................................................................................................1 1.1 Peroxisomes.................................................................................................................. 1 1.1.1 Peroxisome structure and features........................................................................1 1.1.2 Peroxisomal metabolic activity ..............................................................................2 1.1.3 Peroxisome biogenesis...........................................................................................7 1.1.4 Pex3, key component of peroxisome biogenesis................................................ 11 1.1.5 Peroxisome related pathologies ........................................................................
    [Show full text]
  • The Human PEX3 Gene Encoding a Peroxisomal Assembly Protein: Genomic Organization, Positional Mapping, and Mutation Analysis in Candidate Phenotypes
    Biochemical and Biophysical Research Communications 268, 704–710 (2000) doi:10.1006/bbrc.2000.2193, available online at http://www.idealibrary.com on The Human PEX3 Gene Encoding a Peroxisomal Assembly Protein: Genomic Organization, Positional Mapping, and Mutation Analysis in Candidate Phenotypes Ania C. Muntau,* Andreas Holzinger,* Peter U. Mayerhofer,* Jutta Ga¨rtner,† Adelbert A. Roscher,*,1 and Stefan Kammerer*,2 *Dr. von Hauner Children’s Hospital, Laboratory of Molecular Biology, Ludwig-Maximilians-University, Lindwurmstrasse 4, 80337 Munich, Germany; and †Department of Pediatrics, Heinrich Heine University Du¨ sseldorf, Moorenstrasse 5, 40225 Du¨ sseldorf, Germany Received January 6, 2000 Peroxisomes are single-membrane-bound organelles In yeasts, the peroxin Pex3p was identified as a perox- present in all eukaryotic cells other than mature eryth- isomal integral membrane protein that presumably rocytes (1). A large variety of metabolic pathways in- plays a role in the early steps of peroxisomal assembly. cluding the production and degradation of hydrogen In humans, defects of peroxins cause peroxisomal bio- peroxide, and many reactions that involve lipids have genesis disorders such as Zellweger syndrome. We pre- been assigned to the peroxisome (2). The biogenesis of viously reported data on the human PEX3 cDNA and its functional peroxisomes including peroxisome prolifer- protein, which in addition to the peroxisomal targeting ation, membrane biogenesis and peroxisomal matrix sequence contains a putative endoplasmic reticulum protein import, requires the interaction of numerous targeting signal. Here we report the genomic organiza- proteins, designated peroxins, which are encoded by tion, sequencing of the putative promoter region, chro- mosomal localization, and physical mapping of the hu- PEX genes (3).
    [Show full text]
  • Two Splice Variants of Human PEX19 Exhibit Distinct Functions in Peroxisomal Assembly
    Biochemical and Biophysical Research Communications 291, 1180–1186 (2002) doi:10.1006/bbrc.2002.6568, available online at http://www.idealibrary.com on Two Splice Variants of Human PEX19 Exhibit Distinct Functions in Peroxisomal Assembly Peter U. Mayerhofer, Tanja Kattenfeld, Adelbert A. Roscher, and Ania C. Muntau1 Dr. v. Hauner Children’s Hospital, Department of Clinical Chemistry and Biochemical Genetics, Ludwig-Maximilians-University Munich, Lindwurmstrasse 4, D-80337 Munich, Germany Received January 31, 2002 cific biological functions of the different predicted do- PEX19 has been shown to play a central role in the mains of the PEX19 protein. © 2002 Elsevier Science (USA) early steps of peroxisomal membrane synthesis. Com- Key Words: peroxisome biogenesis disorders; splice putational database analysis of the PEX19 sequence variants; farnesylation; peroxin; PEX; ABC half trans- revealed three different conserved domains: D1 (aa porter; peroxisomal membrane proteins. 1–87), D2 (aa 88–272), and D3 (aa 273–299). However, these domains have not yet been linked to specific biological functions. We elected to functionally char- The biogenesis of peroxisomes (reviewed by 1–3) is a acterize the proteins derived from two naturally oc- curring PEX19 splice variants: PEX19⌬E2 lacking the complex process including the biosynthesis of the per- N-terminal domain D1 and PEX19⌬E8 lacking the do- oxisomal membrane, the targeting and insertion of per- main D3. Both interact with peroxisomal ABC trans- oxisomal membrane proteins (PMPs) into this mem- porters (ALDP, ALDRP, PMP70) and with full-length brane, and the import of peroxisomal matrix enzymes PEX3 as shown by in vitro protein interaction studies.
    [Show full text]
  • Gnomad Lof Supplement
    1 gnomAD supplement gnomAD supplement 1 Data processing 4 Alignment and read processing 4 Variant Calling 4 Coverage information 5 Data processing 5 Sample QC 7 Hard filters 7 Supplementary Table 1 | Sample counts before and after hard and release filters 8 Supplementary Table 2 | Counts by data type and hard filter 9 Platform imputation for exomes 9 Supplementary Table 3 | Exome platform assignments 10 Supplementary Table 4 | Confusion matrix for exome samples with Known platform labels 11 Relatedness filters 11 Supplementary Table 5 | Pair counts by degree of relatedness 12 Supplementary Table 6 | Sample counts by relatedness status 13 Population and subpopulation inference 13 Supplementary Figure 1 | Continental ancestry principal components. 14 Supplementary Table 7 | Population and subpopulation counts 16 Population- and platform-specific filters 16 Supplementary Table 8 | Summary of outliers per population and platform grouping 17 Finalizing samples in the gnomAD v2.1 release 18 Supplementary Table 9 | Sample counts by filtering stage 18 Supplementary Table 10 | Sample counts for genomes and exomes in gnomAD subsets 19 Variant QC 20 Hard filters 20 Random Forest model 20 Features 21 Supplementary Table 11 | Features used in final random forest model 21 Training 22 Supplementary Table 12 | Random forest training examples 22 Evaluation and threshold selection 22 Final variant counts 24 Supplementary Table 13 | Variant counts by filtering status 25 Comparison of whole-exome and whole-genome coverage in coding regions 25 Variant annotation 30 Frequency and context annotation 30 2 Functional annotation 31 Supplementary Table 14 | Variants observed by category in 125,748 exomes 32 Supplementary Figure 5 | Percent observed by methylation.
    [Show full text]
  • Structural and Functional Roles of Ether Lipids John M
    Washington University School of Medicine Digital Commons@Becker Open Access Publications 2018 Structural and functional roles of ether lipids John M. Dean Irfan J. Lodhi Follow this and additional works at: https://digitalcommons.wustl.edu/open_access_pubs Protein Cell 2018, 9(2):196–206 DOI 10.1007/s13238-017-0423-5 Protein & Cell REVIEW Structural and functional roles of ether lipids John M. Dean, Irfan J. Lodhi& Division of Endocrinology, Metabolism and Lipid Research, Department of Medicine, Washington University School of Medicine, Saint Louis, MO 63110, USA & Correspondence: [email protected] (I. J. Lodhi) Received March 15, 2017 Accepted April 25, 2017 ABSTRACT found in the brain, heart, spleen, and white blood cells, while Cell liver has scant amount of intracellular ether lipids (Braver- Ether lipids, such as plasmalogens, are peroxisome- & man and Moser, 2012). derived glycerophospholipids in which the hydrocarbon Plasmalogens are the most common form of ether lipids chain at the sn-1 position of the glycerol backbone is and are characterized by a cis double bond adjacent to the attached by an ether bond, as opposed to an ester bond ether linkage. Plasmalogens were serendipitously discov- in the more common diacyl phospholipids. This seem- ered in 1924 by Feulgen and Voit while staining tissue sec- ingly simple biochemical change has profound struc- Protein tions with a nuclear stain that reacts with aldehydes released tural and functional implications. Notably, the tendency by acid hydrolysis of DNA (Snyder, 1999). Because the acid of ether lipids to form non-lamellar inverted hexagonal treatment also resulted in breakdown of the vinyl ether bond structures in model membranes suggests that they have of plasmalogens to generate aldehydes, the researchers a role in facilitating membrane fusion processes.
    [Show full text]
  • Novel Skin Phenotypes Revealed by a Genome-Wide Mouse Reverse Genetic Screen
    ARTICLE Received 9 Jan 2014 | Accepted 4 Mar 2014 | Published 11 Apr 2014 DOI: 10.1038/ncomms4540 OPEN Novel skin phenotypes revealed by a genome-wide mouse reverse genetic screen Kifayathullah Liakath-Ali1,2,3, Valerie E. Vancollie4, Emma Heath1, Damian P. Smedley4, Jeanne Estabel4, David Sunter4, Tia DiTommaso5,w, Jacqueline K. White4, Ramiro Ramirez-Solis4, Ian Smyth5, Karen P. Steel4,6 & Fiona M. Watt1 Permanent stop-and-shop large-scale mouse mutant resources provide an excellent platform to decipher tissue phenogenomics. Here we analyse skin from 538 knockout mouse mutants generated by the Sanger Institute Mouse Genetics Project. We optimize immunolabelling of tail epidermal wholemounts to allow systematic annotation of hair follicle, sebaceous gland and interfollicular epidermal abnormalities using ontology terms from the Mammalian Phenotype Ontology. Of the 50 mutants with an epidermal phenotype, 9 map to human genetic conditions with skin abnormalities. Some mutant genes are expressed in the skin, whereas others are not, indicating systemic effects. One phenotype is affected by diet and several are incompletely penetrant. In-depth analysis of three mutants, Krt76, Myo5a (a model of human Griscelli syndrome) and Mysm1, provides validation of the screen. Our study is the first large-scale genome-wide tissue phenotype screen from the International Knockout Mouse Consortium and provides an open access resource for the scientific community. 1 Centre for Stem Cells and Regenerative Medicine, King’s College London, Guy’s Hospital, London SE1 9RT, UK. 2 Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QW, UK. 3 Wellcome Trust—Medical Research Council Stem Cell Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK.
    [Show full text]
  • Association Between the Intrinsically Disordered Protein PEX19 and PEX3
    Association between the Intrinsically Disordered Protein PEX19 and PEX3 Katarina Hattula., Daniel Hirschberg., Nisse Kalkkinen, Sarah J. Butcher, Ari Ora¤* Institute of Biotechnology, University of Helsinki, Helsinki, Finland Abstract In peroxisomes, peroxins (PEXs) 3 and 19 are the principal protein components of the machinery required for early peroxisomal biogenesis. For further insight into the interaction of PEX3 and PEX19, we used hydrogen exchange mass spectrometry to monitor conformational changes during complex formation between PEX3 and PEX19 in vitro. Our data showed that PEX19 remained highly flexible during interaction with PEX3. However, we could detect three changes, one each in the N-and C-terminus along with a small stretch in the middle of PEX19 (F64–L74) which became shielded from hydrogen exchange when interacting with PEX3. PEX3 became more protected from hydrogen exchange in the binding groove for PEX19 with only small changes elsewhere. Most likely the N-terminus of PEX19 initiates the binding to PEX3, and then subtle conformational changes in PEX3 affect the surface of the PEX3 molecule. PEX19 in turn, is stabilized by folding of a short helix and its C-terminal folding core permitting PEX19 to bind to PEX3 with higher affinity than just the N-terminal interaction allows. Thus within the cell, PEX3 is stabilized by PEX19 preventing PEX3 aggregation. Citation: Hattula K, Hirschberg D, Kalkkinen N, Butcher SJ, Ora A (2014) Association between the Intrinsically Disordered Protein PEX19 and PEX3. PLoS ONE 9(7): e103101. doi:10.1371/journal.pone.0103101 Editor: Elena Papaleo, University of Copenhagen, Denmark Received April 7, 2014; Accepted June 24, 2014; Published July 25, 2014 Copyright: ß 2014 Hattula et al.
    [Show full text]
  • Blueprint Genetics Peroxisomal Disorders Panel
    Peroxisomal Disorders Panel Test code: ME0401 Is a 27 gene panel that includes assessment of non-coding variants. Is ideal for patients with a clinical suspicion of infantile Refsum disease, neonatal adrenoleukodystrophy, rhizomelic chondrodysplasia punctata or Zellweger syndrome. The genes on this panel are included in the Comprehensive Metabolism Panel. About Peroxisomal Disorders Peroxisome biogenesis disorders (PBDs) include a disease continuum of Zellweger syndrome spectrum. This spectrum includes three similar disorders. These are the most severe Zellweger syndrome (ZS), adrenoleukodystrophy with neonatal onset (N-ALD) and the mildest infantile Refsum disease (IRD). Disease onset is typically during childhood, when affected children are hypotonic, have seizures, hepatic dysfunction and distinctive craniofacial dysmorphism. Babies with ZS typically have no developmental progress and die during the first year of life. N-ALD and IRD have more variable manifestations from mild to severe. In addition to the Zellweger syndrome spectrum, PBDs include rhizomelic chondrodysplasia punctata (RCDP). PBDs are caused by mutations in the genes encoding proteins important for peroxisome function, assembly or biogenesis. Mutations in these genes results in the accumulation of very long chain fatty acids and branched chain fatty acids. The prevalence of Zellweger syndrome spectrum disorders is estimated at 1:50 000. In addition to peroxisomal biogenesis disorders, this panel has the ability to diagnose several related peroxisomal disorders such as
    [Show full text]
  • Mouse Screen Reveals Multiple New Genes Underlying Mouse and Human Hearing Loss
    King’s Research Portal DOI: 10.1371/journal.pbio.3000194 Document Version Publisher's PDF, also known as Version of record Link to publication record in King's Research Portal Citation for published version (APA): Ingham, N. J., Pearson, S. A., Vancollie, V. E., Lewis, M. A., Chen, J., Buniello, A., Martelletti, E., & Steel, K. (2019). Mouse screen reveals multiple new genes underlying mouse and human hearing loss. PLoS biology, 17(4), [e3000194]. https://doi.org/10.1371/journal.pbio.3000194 Citing this paper Please note that where the full-text provided on King's Research Portal is the Author Accepted Manuscript or Post-Print version this may differ from the final Published version. If citing, it is advised that you check and use the publisher's definitive version for pagination, volume/issue, and date of publication details. And where the final published version is provided on the Research Portal, if citing you are again advised to check the publisher's website for any subsequent corrections. General rights Copyright and moral rights for the publications made accessible in the Research Portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognize and abide by the legal requirements associated with these rights. •Users may download and print one copy of any publication from the Research Portal for the purpose of private study or research. •You may not further distribute the material or use it for any profit-making activity or commercial gain •You may freely distribute the URL identifying the publication in the Research Portal Take down policy If you believe that this document breaches copyright please contact [email protected] providing details, and we will remove access to the work immediately and investigate your claim.
    [Show full text]
  • Peroxisome Quality Control and Dysregulated Lipid Metabolism in Neurodegenerative Diseases Doo Sin Jo1,Nayeonpark2 and Dong-Hyung Cho1,2
    Jo et al. Experimental & Molecular Medicine (2020) 52:1486–1495 https://doi.org/10.1038/s12276-020-00503-9 Experimental & Molecular Medicine REVIEW ARTICLE Open Access Peroxisome quality control and dysregulated lipid metabolism in neurodegenerative diseases Doo Sin Jo1,NaYeonPark2 and Dong-Hyung Cho1,2 Abstract In recent decades, the role of the peroxisome in physiology and disease conditions has become increasingly important. Together with the mitochondria and other cellular organelles, peroxisomes support key metabolic platforms for the oxidation of various fatty acids and regulate redox conditions. In addition, peroxisomes contribute to the biosynthesis of essential lipid molecules, such as bile acid, cholesterol, docosahexaenoic acid, and plasmalogen. Therefore, the quality control mechanisms that regulate peroxisome biogenesis and degradation are important for cellular homeostasis. Current evidence indicates that peroxisomal function is often reduced or dysregulated in various human disease conditions, such as neurodegenerative diseases. Here, we review the recent progress that has been made toward understanding the quality control systems that regulate peroxisomes and their pathological implications. Introduction for the innate immune response and inflammatory sig- Peroxisomes contain hydrogen peroxide-generating naling4,5. Unsurprisingly, peroxisome dysregulation has fi 1234567890():,; 1234567890():,; 1234567890():,; 1234567890():,; oxidases and were rst described by Christian De Duve been associated with various human disorders, such as in the 1960s1,2. Peroxisomes are tiny (0.2–1-μm-dia- metabolic diseases, cancer, and neurodegenerative dis- meter), single-membrane-bound organelles, and hun- eases, as well as peroxisome biogenesis disorders (PBDs)2. dreds of peroxisomes can exist in a single mammalian Zellweger syndrome, one of the three PBDs, is a rare cell2.
    [Show full text]