Mouse Serpinb7 Conditional Knockout Project (CRISPR/Cas9)
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Characterisation of Serpinb2 As a Stress Response Modulator
University of Wollongong Research Online University of Wollongong Thesis Collection 1954-2016 University of Wollongong Thesis Collections 2015 Characterisation of SerpinB2 as a stress response modulator Jodi Anne Lee University of Wollongong Follow this and additional works at: https://ro.uow.edu.au/theses University of Wollongong Copyright Warning You may print or download ONE copy of this document for the purpose of your own research or study. The University does not authorise you to copy, communicate or otherwise make available electronically to any other person any copyright material contained on this site. You are reminded of the following: This work is copyright. Apart from any use permitted under the Copyright Act 1968, no part of this work may be reproduced by any process, nor may any other exclusive right be exercised, without the permission of the author. Copyright owners are entitled to take legal action against persons who infringe their copyright. A reproduction of material that is protected by copyright may be a copyright infringement. A court may impose penalties and award damages in relation to offences and infringements relating to copyright material. Higher penalties may apply, and higher damages may be awarded, for offences and infringements involving the conversion of material into digital or electronic form. Unless otherwise indicated, the views expressed in this thesis are those of the author and do not necessarily represent the views of the University of Wollongong. Recommended Citation Lee, Jodi Anne, Characterisation of SerpinB2 as a stress response modulator, Doctor of Philosophy thesis, School of Biological Sciences, University of Wollongong, 2015. https://ro.uow.edu.au/theses/4538 Research Online is the open access institutional repository for the University of Wollongong. -
Adaptive Evolution and Divergence of SERPINB3: a Young Duplicate in Great Apes
Adaptive Evolution and Divergence of SERPINB3: A Young Duplicate in Great Apes Sı´lvia Gomes1*, Patrı´cia I. Marques1,2, Rune Matthiesen3, Susana Seixas1* 1 Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Porto, Portugal, 2 Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Porto, Portugal, 3 National Health Institute Doutor Ricardo Jorge (INSA), Lisboa, Portugal Abstract A series of duplication events led to an expansion of clade B Serine Protease Inhibitors (SERPIN), currently displaying a large repertoire of functions in vertebrates. Accordingly, the recent duplicates SERPINB3 and B4 located in human 18q21.3 SERPIN cluster control the activity of different cysteine and serine proteases, respectively. Here, we aim to assess SERPINB3 and B4 coevolution with their target proteases in order to understand the evolutionary forces shaping the accelerated divergence of these duplicates. Phylogenetic analysis of primate sequences placed the duplication event in a Hominoidae ancestor (,30 Mya) and the emergence of SERPINB3 in Homininae (,9 Mya). We detected evidence of strong positive selection throughout SERPINB4/B3 primate tree and target proteases, cathepsin L2 (CTSL2) and G (CTSG) and chymase (CMA1). Specifically, in the Homininae clade a perfect match was observed between the adaptive evolution of SERPINB3 and cathepsin S (CTSS) and most of sites under positive selection were located at the inhibitor/protease interface. Altogether our results seem to favour a coevolution hypothesis for SERPINB3, CTSS and CTSL2 and for SERPINB4 and CTSG and CMA1.A scenario of an accelerated evolution driven by host-pathogen interactions is also possible since SERPINB3/B4 are potent inhibitors of exogenous proteases, released by infectious agents. -
Mutations in SERPINB7, Encoding a Member of the Serine Protease Inhibitor Superfamily, Cause Nagashima-Type Palmoplantar Keratosis
REPORT Mutations in SERPINB7, Encoding a Member of the Serine Protease Inhibitor Superfamily, Cause Nagashima-type Palmoplantar Keratosis Akiharu Kubo,1,2,3,* Aiko Shiohama,1,4 Takashi Sasaki,1,2,3 Kazuhiko Nakabayashi,5 Hiroshi Kawasaki,1 Toru Atsugi,1,6 Showbu Sato,1 Atsushi Shimizu,7 Shuji Mikami,8 Hideaki Tanizaki,9 Masaki Uchiyama,10 Tatsuo Maeda,10 Taisuke Ito,11 Jun-ichi Sakabe,11 Toshio Heike,12 Torayuki Okuyama,13 Rika Kosaki,14 Kenjiro Kosaki,15 Jun Kudoh,16 Kenichiro Hata,5 Akihiro Umezawa,17 Yoshiki Tokura,11 Akira Ishiko,18 Hironori Niizeki,19 Kenji Kabashima,9 Yoshihiko Mitsuhashi,10 and Masayuki Amagai1,2,4 ‘‘Nagashima-type’’ palmoplantar keratosis (NPPK) is an autosomal recessive nonsyndromic diffuse palmoplantar keratosis characterized by well-demarcated diffuse hyperkeratosis with redness, expanding on to the dorsal surfaces of the palms and feet and the Achilles tendon area. Hyperkeratosis in NPPK is mild and nonprogressive, differentiating NPPK clinically from Mal de Meleda. We performed whole-exome and/or Sanger sequencing analyses of 13 unrelated NPPK individuals and identified biallelic putative loss-of-function mutations in SERPINB7, which encodes a cytoplasmic member of the serine protease inhibitor superfamily. We identified a major caus- ative mutation of c.796C>T (p.Arg266*) as a founder mutation in Japanese and Chinese populations. SERPINB7 was specifically present in the cytoplasm of the stratum granulosum and the stratum corneum (SC) of the epidermis. All of the identified mutants are predicted to cause premature termination upstream of the reactive site, which inhibits the proteases, suggesting a complete loss of the protease inhibitory activity of SERPINB7 in NPPK skin. -
Serpins—From Trap to Treatment
MINI REVIEW published: 12 February 2019 doi: 10.3389/fmed.2019.00025 SERPINs—From Trap to Treatment Wariya Sanrattana, Coen Maas and Steven de Maat* Department of Clinical Chemistry and Haematology, University Medical Center Utrecht, Utrecht University, Utrecht, Netherlands Excessive enzyme activity often has pathological consequences. This for example is the case in thrombosis and hereditary angioedema, where serine proteases of the coagulation system and kallikrein-kinin system are excessively active. Serine proteases are controlled by SERPINs (serine protease inhibitors). We here describe the basic biochemical mechanisms behind SERPIN activity and identify key determinants that influence their function. We explore the clinical phenotypes of several SERPIN deficiencies and review studies where SERPINs are being used beyond replacement therapy. Excitingly, rare human SERPIN mutations have led us and others to believe that it is possible to refine SERPINs toward desired behavior for the treatment of enzyme-driven pathology. Keywords: SERPIN (serine proteinase inhibitor), protein engineering, bradykinin (BK), hemostasis, therapy Edited by: Marvin T. Nieman, Case Western Reserve University, United States INTRODUCTION Reviewed by: Serine proteases are the “workhorses” of the human body. This enzyme family is conserved Daniel A. Lawrence, throughout evolution. There are 1,121 putative proteases in the human body, and about 180 of University of Michigan, United States Thomas Renne, these are serine proteases (1, 2). They are involved in diverse physiological processes, ranging from University Medical Center blood coagulation, fibrinolysis, and inflammation to immunity (Figure 1A). The activity of serine Hamburg-Eppendorf, Germany proteases is amongst others regulated by a dedicated class of inhibitory proteins called SERPINs Paulo Antonio De Souza Mourão, (serine protease inhibitors). -
Suppression of the Invasion and Migration of Cancer Cells by SERPINB Family Genes and Their Derived Peptides
238 ONCOLOGY REPORTS 27: 238-245, 2012 Suppression of the invasion and migration of cancer cells by SERPINB family genes and their derived peptides RUEY-HWANG CHOU1-4, HUI-CHIN WEN1,7, WEI-GUANG LIANG1,5, SHENG-CHIEH LIN1,5, HSIAO-WEI YUAN1, CHENG-WEN WU1,5,6 and WUN-SHAING WAYNE CHANG1 1National Institute of Cancer Research, National Health Research Institutes, Miaoli 35053; 2Center for Molecular Medicine, China Medical University Hospital, Taichung 40402; 3China Medical University, Taichung 40402; 4Department of Biotechnology, Asia University, Taichung 41354; 5College of Life Science, National Tsing Hua University, Hsinchu 30013; 6Institute of Biochemistry and Molecular Biology, National Yang Ming University, Taipei 11221, Taiwan, R.O.C. Received June 28, 2011; Accepted August 17, 2011 DOI: 10.3892/or.2011.1497 Abstract. Apart from SERPINB2 and SERPINB5, the roles SERPINB RCL-peptides may provide a reasonable strategy of the remaining 13 members of the human SERPINB family against lethal cancer metastasis. in cancer metastasis are still unknown. In the present study, we demonstrated that most of these genes are differentially Introduction expressed in tumor tissues compared to matched normal tissues from lung or breast cancer patients. Overexpression of Cancer metastasis is the leading cause of morbidity and each SERPINB gene effectively suppressed the invasiveness mortality in cancer patients. It is a highly complex process, and motility of malignant cancer cells. Among all of the genes, including cell detachment, migration, invasion, circulation in the SERPINB1, SERPINB5 and SERPINB7 genes were more blood vessels, adhesion, colonization at other sites and forma- potent, and the inhibitory effect was further enhanced by tion of secondary tumors (1). -
Correlation of Serpin–Protease Expression by Comparative Analysis of Real-Time PCR Profiling Data
View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Genomics 88 (2006) 173–184 www.elsevier.com/locate/ygeno Correlation of serpin–protease expression by comparative analysis of real-time PCR profiling data Sunita Badola a, Heidi Spurling a, Keith Robison a, Eric R. Fedyk a, Gary A. Silverman b, ⁎ Jochen Strayle c, Rosana Kapeller a,1, Christopher A. Tsu a, a Millennium Pharmaceuticals, Inc., 40 Landsdowne Street, Cambridge, MA 02139, USA b Department of Pediatrics, University of Pittsburgh School of Medicine, Magee-Women’s Hospital, 300 Halket Street, Pittsburgh, PA 15213, USA c Bayer HealthCare AG, 42096 Wuppertal, Germany Received 2 December 2005; accepted 27 March 2006 Available online 18 May 2006 Abstract Imbalanced protease activity has long been recognized in the progression of disease states such as cancer and inflammation. Serpins, the largest family of endogenous protease inhibitors, target a wide variety of serine and cysteine proteases and play a role in a number of physiological and pathological states. The expression profiles of 20 serpins and 105 serine and cysteine proteases were determined across a panel of normal and diseased human tissues. In general, expression of serpins was highly restricted in both normal and diseased tissues, suggesting defined physiological roles for these protease inhibitors. A high correlation in expression for a particular serpin–protease pair in healthy tissues was often predictive of a biological interaction. The most striking finding was the dramatic change observed in the regulation of expression between proteases and their cognate inhibitors in diseased tissues. -
The Aggregation-Prone Intracellular Serpin SRP-2 Fails to Transit the ER in Caenorhabditis Elegans
GENETICS | INVESTIGATION The Aggregation-Prone Intracellular Serpin SRP-2 Fails to Transit the ER in Caenorhabditis elegans Richard M. Silverman, Erin E. Cummings, Linda P. O’Reilly, Mark T. Miedel, Gary A. Silverman, Cliff J. Luke, David H. Perlmutter, and Stephen C. Pak1 Departments of Pediatrics and Cell Biology, University of Pittsburgh School of Medicine, Children’s Hospital of Pittsburgh of University of Pittsburgh Medical Center and Magee–Womens Hospital Research Institute, Pittsburgh, Pennsylvania 15224 ABSTRACT Familial encephalopathy with neuroserpin inclusions bodies (FENIB) is a serpinopathy that induces a rare form of presenile dementia. Neuroserpin contains a classical signal peptide and like all extracellular serine proteinase inhibitors (serpins) is secreted via the endoplasmic reticulum (ER)–Golgi pathway. The disease phenotype is due to gain-of-function missense mutations that cause neuroserpin to misfold and aggregate within the ER. In a previous study, nematodes expressing a homologous mutation in the endogenous Caenorhabditis elegans serpin, srp-2,werereportedtomodeltheERproteotoxicityinducedbyanallele of mutant neuroserpin. Our results suggest that SRP-2 lacksaclassicalN-terminalsignalpeptideandisamemberofthe intracellular serpin family. Using confocal imaging and an ER colocalization marker, we confirmed that GFP-tagged wild-type SRP-2 localized to the cytosol and not the ER. Similarly, the aggregation- prone SRP-2 mutant formed intracellular inclusions that localized to the cytosol. Interestingly, wild-type SRP-2,targetedtotheERbyfusion to a cleavable N-terminal signal peptide, failedtobesecretedandaccumulatedwithintheERlumen.ThisERretentionphenotypeistypical of other obligate intracellular serpins forced to translocate across the ER membrane. Neuroserpin is a secreted protein that inhibits trypsin- like proteinase. SRP-2 is a cytosolic serpin that inhibits lysosomal cysteine peptidases. We concluded that SRP-2 is neither an ortholog nor a functional homolog of neuroserpin. -
Targeting the IL33–NLRP3 Axis Improves Therapy for Experimental Cerebral Malaria
Targeting the IL33–NLRP3 axis improves therapy for experimental cerebral malaria Patrick Strangwarda, Michael J. Haleya,1, Manuel G. Albornoza,1, Jack Barringtona,1, Tovah Shawa, Rebecca Dookiea, Leo Zeefa, Syed M. Bakera, Emma Wintera, Te-Chen Tzengb, Douglas T. Golenbockb, Sheena M. Cruickshanka, Stuart M. Allana, Alister Craigc, Foo Y. Liewd,e, David Brougha,2,3, and Kevin N. Coupera,2,3 aSchool of Biological Sciences, Faculty of Biology, Medicine, and Health, Manchester Academic Health Science Centre, University of Manchester, Manchester M13 9PT, United Kingdom; bDivision of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester, MA 01605; cDepartment of Parasitology, Liverpool School of Tropical Medicine, Liverpool L3 5QA, United Kingdom; dDepartment of Immunology, Institute of Infection, Immunity, and Inflammation, University of Glasgow, Glasgow G12 8TA, United Kingdom; and eSchool of Biology and Basic Medical Sciences, Soochow University, 215006 Suzhou, China Edited by Michael B. A. Oldstone, The Scripps Research Institute, La Jolla, CA, and approved June 4, 2018 (received for review January 30, 2018) Cerebral malaria (CM) is a serious neurological complication caused recovery by activating the brain endothelium, causing permeability by Plasmodium falciparum infection. Currently, the only treatment of the blood–brain barrier, activation of astrocytes and microglia, for CM is the provision of antimalarial drugs; however, such treat- disruption of neuronal signaling, and recruitment of circulating ment by itself often fails to prevent death or development of neu- leukocytes (1, 7–9). All of these events have been observed in rological sequelae. To identify potential improved treatments for brains of individuals with fatal CM (1, 6–9). -
Understanding the Role of Plasminogen Activator Inhibitor Type-2 (PAI-2, Serpinb2)
University of Wollongong Research Online University of Wollongong Thesis Collection University of Wollongong Thesis Collections 2011 Understanding the role of plasminogen activator inhibitor type-2 (PAI-2, SerpinB2) in cancer: the relationship between biochemical properties and cellular function Blake J. Cochran University of Wollongong Recommended Citation Cochran, Blake J., Understanding the role of plasminogen activator inhibitor type-2 (PAI-2, SerpinB2) in cancer: the relationship between biochemical properties and cellular function, Doctor of Philosophy thesis, University of Wollongong. School of Biological Sciences, University of Wollongong, 2011. http://ro.uow.edu.au/theses/3300 Research Online is the open access institutional repository for the University of Wollongong. For further information contact Manager Repository Services: [email protected]. UNDERSTANDING THE ROLE OF PLASMINOGEN ACTIVATOR INHIBITOR TYPE-2 (PAI-2, SERPINB2) IN CANCER: THE RELATIONSHIP BETWEEN BIOCHEMICAL PROPERTIES AND CELLULAR FUNCTION A thesis submitted in fulfilment of the requirements for the award of the degree Doctor of Philosophy from University of Wollongong by Blake J. Cochran Bachelor of Biotechnology (Honours 1)(Advanced) School of Biological Sciences University of Wollongong 2011 CERTIFICATION I, Blake J. Cochran, declare that this thesis, submitted in fulfilment of the requirements for the award of Doctor of Philosophy, in the School of Biological Sciences, University of Wollongong, is wholly my own work unless otherwise referenced or acknowledged. The document has not been submitted for qualifications at any other academic institution. Blake J. Cochran 18 July 2011 I LIST OF PUBLICATIONS Cochran, B.J., Gunawardhana, L.P., Vine, K.L., Lee, J.A., Lobov, S. and Ranson, M. 2009. -
Nagashima-Type Palmoplantar Keratosis in a Chinese Han Population
MOLECULAR MEDICINE REPORTS 14: 4049-4054, 2016 Nagashima-type palmoplantar keratosis in a Chinese Han population JIA ZHANG1*, GUOLONG ZHANG2*, CHENG NI1, RUHONG CHENG1, JIANYING LIANG1, MING LI1 and ZHIRONG YAO1 1Department of Dermatology, Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai 200092; 2Department of Phototherapy, Shanghai Skin Disease Hospital, Shanghai 200050, P.R. China Received July 11, 2015; Accepted May 13, 2016 DOI: 10.3892/mmr.2016.5757 Abstract. Nagashima-type palmoplantar keratosis (NPPK) is an Introduction autosomal recessive form of palmoplantar keratoderma (PPK), which is caused by mutations in the SERPINB7 gene. NPPK Nagashima-type palmoplantar keratosis (NPPK; has only been reported in Japanese and Chinese populations. OMIM #615598) is an autosomal recessive form of palmo- The present study was conducted on 12 unrelated Chinese plantar keratoderma (PPK), which exhibits a relatively high patients who were clinically predicted to suffer from NPPK. incidence and has only been reported in Japanese and Chinese Mutation screening was performed by direct sequencing of the populations (1,2). The characteristic features of NPPK are entire coding regions of SERPINB7, SLURP1, AQP5, CSTA, erythema and hyperkeratosis of the palms and soles, with KRT1 and KRT9 genes. Direct sequencing of SERPINB7 sharp demarcation that mainly extends to the wrists, ankles, revealed five homozygous founder mutations (c.796C>T) Achilles tendon area, and the dorsal aspects of the fingers and and four compound heterozygous mutations in nine patients, toes (1). Furthermore, other frictional regions, such as knees including one novel mutation (c.122_127delTGGTCC). Nine and elbows, may be involved. Some patients with NPPK also out of the 12 patients were diagnosed with NPPK due to exhibit associated palmoplantar features, including a white, SERPINB7 pathogenic mutations, and the results expanded spongy appearance within 10 min of water exposure, hyperhi- the known mutation spectrum of NPPK. -
Hepatic Fibrosis and Carcinogenesis in A1-Antitrypsin Deficiency: A
Downloaded from http://cshperspectives.cshlp.org/ on September 27, 2021 - Published by Cold Spring Harbor Laboratory Press Hepatic Fibrosis and Carcinogenesis in a1-Antitrypsin Deficiency: A Prototype for Chronic Tissue Damage in Gain-of-Function Disorders David H. Perlmutter and Gary A. Silverman Departments of Pediatrics, Cell Biology, and Physiology, University of Pittsburgh School of Medicine, Children’s Hospital of Pittsburgh and Magee-Womens Hospital of UPMC, Pittsburgh, Pennsylvania 15224 Correspondence: [email protected] In a1-antitrypsin (AT) deficiency, a point mutation renders a hepatic secretory glycoprotein prone to misfolding and polymerization. The mutant protein accumulates in the endoplas- mic reticulum of liver cells and causes hepatic fibrosis and hepatocellular carcinoma by a gain-of-function mechanism. Genetic and/or environmental modifiers determine whether an affected homozygote is susceptible to hepatic fibrosis/carcinoma. Two types of proteo- stasis mechanisms for such modifiers have been postulated: variation in the function of intracellular degradative mechanisms and/or variation in the signal transduction pathways that are activated to protect the cell from protein mislocalization and/or aggregation. In recent studies we found that carbamazepine, a drug that has been used safely as an anti- convulsant and mood stabilizer, reduces the hepatic load of mutant ATand hepatic fibrosis in a mouse model by enhancing autophagic disposal of this mutant protein. These results provide evidence that pharmacological manipulation of endogenous proteostasis mechan- isms is an appealing strategy for chemoprophylaxis in disorders involving gain-of-function mechanisms. he classical form of a1-antitrypsin (AT) in 1963, we now know that this liver disease Tdeficiency is a relatively common genetic results from the accumulation of mutant AT disease that causes hepatic fibrosis and carcino- inside of liver cells. -
Nucleotide Variations in Genes Encoding Plasminogen Activator Inhibitor-2 and Serine Proteinase Inhibitor B10 Associated with Prostate Cancer
J Hum Genet (2005) 50: 507–515 DOI 10.1007/s10038-005-0285-1 ORIGINAL ARTICLE Go Shioji Æ Yoichi Ezura Æ Toshiaki Nakajima Kenji Ohgaki Æ Hiromichi Fujiwara Æ Yoshinobu Kubota Tomohiko Ichikawa Æ Katsuki Inoue Æ Taro Shuin Tomonori Habuchi Æ Osamu Ogawa Æ Taiji Nishimura Mitsuru Emi Nucleotide variations in genes encoding plasminogen activator inhibitor-2 and serine proteinase inhibitor B10 associated with prostate cancer Received: 2 May 2005 / Accepted: 25 July 2005 / Published online: 20 September 2005 Ó The Japan Society of Human Genetics and Springer-Verlag 2005 Abstract Genes encoding the serine proteinase inhibitor variations within SERPINB loci at 18q21 might be Bfamily(SERPINBs) are mainly clustered on human associated with risk of prostate cancer in Japanese men. chromosome 18 (18q21). Several serpins are known to A case-control study involving 292 prostate-cancer pa- affect malignant phenotypes of tumor cells, so aberrant tients and 384 controls revealed significant differences in genetic variants in this molecular family are candidates regard to distribution of four missense variations in genes for conferring susceptibility for risk of cancer. We encoding plasminogen activator inhibitor 2 (PAI2)and investigated whether eight selected non-synonymous SERPINB10. The most significant association was de- tected for the N120D polymorphism in the PAI2 gene À5 G. Shioji Æ Y. Ezura Æ T. Nakajima Æ K. Ohgaki (P=5.0·10 ); men carrying the 120-N allele (120-N/N H. Fujiwara Æ M. Emi and 120-N/D genotypes) carried a 2.4-fold increased risk Department of Molecular Biology, Institute of Gerontology, of prostate cancer (95% confidence interval 1.45–4.07).