Heterologous Expression of the Alba Protein from the Hyperthermophilic Archaeon Aeropyrum Pernix

被引:2
|
作者
Crnigoj, Miha [1 ]
Hanzlowsky, Andrej [1 ]
Vilfan, Tanja [1 ]
Ulrih, Natasa Poklar [1 ,2 ]
机构
[1] Univ Ljubljana, Biotech Fac, Dept Food Sci & Technol, Ljubljana 1000, Slovenia
[2] CipKeBiP Ctr Excellence Integrated Approaches Che, Ljubljana 1000, Slovenia
关键词
Aeropyrum pernix; hyperthermophile; archaea; Alba; DNA binding; DNA stabilisation; DNA-BINDING PROTEIN; X-RAY ANALYSIS; CHROMATIN PROTEIN; CRYSTALLIZATION;
D O I
10.5562/cca1772
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Nucleic acid binding proteins have important roles in DNA and RNA packaging, stabilisation and repair, and in gene regulation, and they are therefore essential for all organisms. All of the known hyperthermophiles have at least one DNA sequence encoding for the Alba proteins. The Alba proteins are small (approximately 10 kDa), DNA-binding, basic proteins that appear to partly compensate for the lack of histones in the archaea Aeropyrum pernix and other hyperthermophiles. Two sequences of these potential histone counterparts, the Alba proteins, were identified in the Aeropyrum pernix genome (APE1832.1 and APE 1823). By using a wide range of experimental techniques and by examining several combinations of expression systems the expression of recombinant Alba 1 and Alba2 proteins was optimized. Co-expression of both of the Alba proteins was needed when isolating recombinant Alba2. The purification of both recombinant Alba 1 and Alba2 His-tagged proteins were simplyfied in satisfactory yield. The electrophoretic mobility shift assay demonstrated the ability of the Alba 1 and Alba2 proteins from Aeropyrum pernix to bind DNA. (doi: 10.5562/cca1772)
引用
收藏
页码:499 / 504
页数:6
相关论文
共 50 条
  • [31] Characterization of thermostable native alkaline phosphatase from an aerobic hyperthermophilic archaeon, Aeropyrum pernix K1
    Helianti, Is
    Okubo, Takako
    Morita, Yasutaka
    Tamiya, Eiichi
    APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2007, 74 (01) : 107 - 112
  • [32] Enhanced Activity and Enantioselectivity of a Hyperthermophilic Esterase from Archaeon Aeropyrum pernix K1 by Acetone Treatment
    Fangdi Cong
    Kezhi Xing
    Renjun Gao
    Shugui Cao
    Guirong Zhang
    Applied Biochemistry and Biotechnology, 2011, 165 : 795 - 801
  • [33] Effect of different fluorescent dyes on thermal stability of DNA and cell viability of the hyperthermophilic archaeon Aeropyrum pernix
    Miha Črnigoj
    Rok Kostanjšek
    Gönül Kaletunç
    Nataša Poklar Ulrih
    World Journal of Microbiology and Biotechnology, 2008, 24 : 2115 - 2123
  • [34] Crystal Structure of Novel Dye-linked L-Proline Dehydrogenase from Hyperthermophilic Archaeon Aeropyrum pernix
    Sakuraba, Haruhiko
    Satomura, Takenori
    Kawakami, Ryushi
    Kim, Kwang
    Hara, Yusuke
    Yoneda, Kazunari
    Ohshima, Toshihisa
    JOURNAL OF BIOLOGICAL CHEMISTRY, 2012, 287 (24) : 20070 - 20080
  • [35] Overexpression, purification and crystallization of tyrosyl-tRNA synthetase from the hyperthermophilic archaeon Aeropyrum pernix K1
    Iwaki, J
    Suzuki, R
    Fujimoto, Z
    Momma, M
    Kuno, A
    Hasegawa, T
    ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, 2005, 61 : 1003 - 1005
  • [36] Stability of diether C25,25 liposomes from the hyperthermophilic archaeon Aeropyrum pernix K1
    Gmajner, Dejan
    Ota, Ajda
    Sentjurc, Marjeta
    Ulrih, Natasa Poklar
    CHEMISTRY AND PHYSICS OF LIPIDS, 2011, 164 (03) : 236 - 245
  • [37] Crystallization and preliminary X-ray diffraction studies of a novel alcohol dehydrogenase from the hyperthermophilic archaeon Aeropyrum pernix
    Guy, JE
    Isupov, MN
    Littlechild, JA
    ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2003, 59 : 174 - 176
  • [38] Gene Expression and Characterization of a Third Type of Dye-Linked L-Proline Dehydrogenase from the Aerobic Hyperthermophilic Archaeon, Aeropyrum pernix
    Satomura, Takenori
    Hara, Yusuke
    Suye, Shin-ichiro
    Sakuraba, Haruhiko
    Ohshima, Toshihisa
    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 2012, 76 (03) : 589 - 593
  • [39] An extremely heat-stable extracellular proteinase (aeropyrolysin) from the hyperthermophilic archaeon Aeropyrum pernix K1
    Sako, Y
    Croocker, PC
    Ishida, Y
    FEBS LETTERS, 1997, 415 (03) : 329 - 334
  • [40] Effect of different fluorescent dyes on thermal stability of DNA and cell viability of the hyperthermophilic archaeon Aeropyrum pernix
    Crnigoj, Miha
    Kostanjsek, Rok
    Katletunc, Goenuel
    Ulrih, Natasa Poklar
    WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY, 2008, 24 (10): : 2115 - 2123