• 中国精品科技期刊
  • 中国高校百佳科技期刊
  • 中国中文核心期刊
  • 中国科学引文数据库核心期刊
Advanced Search
HONG Liu, LANG Junchao, HE Dongmei, LIU Kunfeng, WU Jie. Studies on preparation and properties of Staphylococcus nuclease[J]. Journal of China Pharmaceutical University, 2017, 48(2): 214-219. DOI: 10.11665/j.issn.1000-5048.20170213
Citation: HONG Liu, LANG Junchao, HE Dongmei, LIU Kunfeng, WU Jie. Studies on preparation and properties of Staphylococcus nuclease[J]. Journal of China Pharmaceutical University, 2017, 48(2): 214-219. DOI: 10.11665/j.issn.1000-5048.20170213
shu

Studies on preparation and properties of Staphylococcus nuclease

More Information
    Graphical Abstract
    • Abstract
  • In order to study the relationship between Staphylococcal nuclease(SNase)and diabetes mellitus, genetic engineering bacteria E. coli BL21/pET28a-His-SNase was constructed, the expression of soluble extracellular protein SNase was induced and the a preliminary research was made on it. An expression vector pET28a-His-SNase plasmid containing the His-SNase gene was constructed and transformed into E. coli BL21(DE3)competent cells. The protein was induced by lactose and purified by ultrasound destruction and Ni-affinity chromatography, respectively. It was then analyzed by SDS-PAGE and Western blot. The enzymatic properties for SNase has been preliminary studied as well. Results indicated that the purity of the correctly expressed fusion protein His-SNase was over 85%. SNase showed good activity within a wide range of pH and good heat resistance. This experiment might be a foundation work for the further study on the relationship between SNase and with diabetes.
    • FullText(HTML)
    • References (23)
  • [1]
    Haskins K,Portas M,Bergman B,et al.Pancreatic islet-specific T-cell clones from nonobese diabetic mice[J].Proc Nat Acad Sci U S A,1989,86(20):8000-8004.
    [2]
    Panina-Bordignon P,Lang R,van Endert PM,et al.Cytotoxic T cells specific for glutamic acid decarboxylase in autoimmune diabetes[J].J Exp Med,1995,181(5):1923-1927.
    [3]
    Bekris LM,Kavanagh TJ,Lernmark A.Targeting type 1 diabetes before and at the clinical onset of disease[J].Endocr Metab Immune Disord Drug Targets,2006,6(1):103-124.
    [4]
    White SA,Shaw JA,Sutherland DER.Pancreas transplantation[J].Lancet,2009,373(9677):1808-1817.
    [5]
    Wen Y,Chen B,Ildsrad ST.Stem cell-based strategies for the treatment of type 1 diabetes mellitus[J].Expert Opin Biol Ther,2011,11(1):41-53.
    [6]
    Mócsai A. Diverse novel functions of neutrophils in immunity,inflammation,and beyond[J].J Exp Med,2013,210(7):1283-1299.
    [7]
    Segal AW.How neutrophils kill microbes[J].Annurev Immunol,2005,23:197-223.
    [8]
    Brinkmann V,R eichard U,Goosmann C,et al.Neutrophil extracellular traps kill bacteria[J].Science,2004,303(5663):1532-1535.
    [9]
    Darrah E,Andrade F.NETs:the missing link between cell death and systemic autoimmune diseases[J].J Front Immunol,2012,3:428.
    [10]
    Villanueva E,Yalavarthi S,Berthier CC,et al.Netting neutrophils induce endothelial damage,infiltrate tissues,and expose immunostimulatory molecules in systemic lupus erythematosus[J].J Immunol,2011,187(1):538-552.
    [11]
    Kessenbrock K,Krumbholz M,Schonermarck U,et al.Netting neutrophils in autoimmune small-vessel vasculitis[J].Nat Med,2009,15(6):623-625.
    [12]
    Khandpur R,Carmona-Rivera C,Vivekanandan-Giri A,et al.NETs are a source of citrullinated autoantigens and stimulate inflammatory responses in rheumatoid arthritis[J].Sci Transl Med,2013,5(178):178ra40.
    [13]
    Wang Y,Xiao Y,Zhong L,et al.Increased neutrophil elastase and proteinase 3 and augmented NETosis are closely associated with β-cell autoimmunity in patients with type 1 diabetes[J].Diabetes,2014,63(12):4239-4248.
    [14]
    Diana J,Simoni Y,Furio L,et al.Crosstalk between neutrophils,B-1a cells and plasmacytoid dendritic cells initiates autoimmune diabetes[J].Nat Med,2013,19(1):65-73.
    [15]
    Dwivedi N,Upadhyay J,Neeli I,et al.Felty′s syndrome autoantibodies bind to deiminated histones and neutrophil extracellular chromatin traps[J].Arthritis Rheum,2012,64(4):982-992.
    [16]
    Nakashima K,Hagiwara T,Yamada M.Nuclear localization of peptidylarginine deiminase V and histone deimination in granulocytes[J].J Biol Chem,2002,277(51):49562-49568.
    [17]
    Simon D,Simon HU,Yousefi S.Extracellular DNA traps in allergic,infectious,and autoimmune diseases[J].Allergy,2013,68:409-416.
    [18]
    Taniuchi H,Anfinsen CB.Simultaneous formation of two alternative enzymology active structures by complementation of two overlapping fragments of staphylococcal nuclease[J].J Biol Chem,1971,246(7):2291-2301.
    [19]
    Shortle D,Meeker AK.Residual structure in large fragments of staphylococcal nuclease:effects of amino acid substitutions[J].Biochemistry,1989,28(3):936-944.
    [20]
    Singh PK,Chan PF,Hibbs MJ,et al.High-yield production and characterization of biologically active GST-tagged human topoisomeraseIIα protein in insect cells for the development of a high-throughput assay[J].Protein Expr Purif,2011,76(2):165-172.
    [21]
    Vorackova I,Suchanova S,Ulbrich P,et al.Purification of proteins containing zinc finger domains using immobilized metal ion affinity chromatography[J].Protein Expr Purif,2011,79(1):88-95.
    [22]
    Zakalskiy AE,Zakalska OM,Rzhepetskyy YA,et al.Overexpression of(His)6 tagged human arginase I in Saccharomyces cerevisiae and enzyme purification using metal affinity chromatography[J].Protein Expr Purif,2012,81(1):63-68.
    [23]
    Shiokawa D,Tanuma S.Characteration of human DNase1 family endonucleases and activation of DNase gamma during apoptosis[J].Bioehemistry,2001,40:143-52.
    • Related Articles

  • Related Articles

    [1]CHEN Xi, JIN Liang. Research progress on sumoylation in type 2 diabetes mellitus[J]. Journal of China Pharmaceutical University, 2025, 56(1): 125-131. DOI: 10.11665/j.issn.1000-5048.2023112002
    [2]MU Jinming, LIU Yue, ZHANG Fangfang, JIN Liang. Relationship between circular RNA and type 2 diabetes and its clinical application[J]. Journal of China Pharmaceutical University, 2020, 51(3): 374-378. DOI: 10.11665/j.issn.1000-5048.20200316
    [3]LI Yongrong, CHENG Tao, WANG Yongsheng, LI Qin, GAO Bo, HE Yun, BAI Yu. Effect of extract of selenium-enriched Astragalus membranaceus on insulin resistance in streptozotocin-induced diabetic rats[J]. Journal of China Pharmaceutical University, 2018, 49(6): 739-745. DOI: 10.11665/j.issn.1000-5048.20180616
    [4]DU Te, CHEN Jing, SHEN Xu. Research advances in susceptibility genes of non-alcoholic fatty liver disease and its association with type 2 diabetes[J]. Journal of China Pharmaceutical University, 2018, 49(5): 537-544. DOI: 10.11665/j.issn.1000-5048.20180504
    [5]CAO Lanjie, ZHU Kuiniu, JIANG Cuihua, GUNN ·Jordan, CHE Chun-Tao, ZHANG Jian, YIN Zhiqi. Intervention effects of the extract of Alismatis Rhizoma on streptozotocin-induced type 2 diabetic rats[J]. Journal of China Pharmaceutical University, 2017, 48(5): 601-608. DOI: 10.11665/j.issn.1000-5048.20170516
    [6]WANG Xinning, XU Bin, ZHOU Jinpei, ZHANG Huibin. Advances of anti-diabetic drugs based on new targets[J]. Journal of China Pharmaceutical University, 2015, 46(2): 141-152. DOI: 10.11665/j.issn.1000-5048.20150202
    [7]LI Chen, YUAN Linhua, MA Xueqin, LIU Xiaoquan, XI Tao. Variation of cardiovascular risk biomarkers in type 2 diabetes mellitus[J]. Journal of China Pharmaceutical University, 2013, 44(3): 257-262. DOI: 10.11665/j.issn.1000-5048.20130314
    [8]HE Bi-xia, QIAO Li-yun, PENG Jun, XIE Zhi-ping, DING Qi-long. Effects and mechanisms of mifepristone on insulin-resistant HepG 2 cells[J]. Journal of China Pharmaceutical University, 2011, 42(2): 153-159.
    [9]Influences on Serum Insulin and Glucose by Different Fasting Time in Alloxan Diabetic Mice[J]. Journal of China Pharmaceutical University, 2001, (3): 59-62.
    [10]Determination of the Concentration of Intracellular Calcium in Lymphocyte by Fluo 3 AM and Study of the Influence of the Drugs on Calcium Release[J]. Journal of China Pharmaceutical University, 1999, (6): 53-57.

Catalog

    Get Citation
    PDF
    XML
    Article views (880) PDF downloads (1457) Cited by()
    Turn off MathJax
    Article Contents
    Abstract
    References
    Related Articles

    Copyright © Journal of China Pharmaceutical University苏ICP备12050101号-2

    Address：24 Tongjia Lane, Nanjing City, Jiangsu Province （210009）Tel: 025-83271566E-mail: xuebao@cpu.edu.cn

    Supported by: Beijing Renhe Information Technology Co., Ltd.  Baidu Analytics

    Total visits：300499

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return