Early steps in the unfolding of thermolysin-like proteases

被引:26
|
作者
Vriend, G
Berendsen, HJC
van den Burg, B
Venema, G
Eijsink, VGH
机构
[1] European Mol Biol Lab, D-69117 Heidelberg, Germany
[2] Univ Groningen, BIOSON Res Inst, NL-9747 AG Groningen, Netherlands
[3] Univ Groningen, Dept Genet, Groningen Biomol Sci & Biotechnol Inst, NL-9751 NN Haren, Netherlands
[4] Agr Univ Norway, Dept Biotechnol Sci, N-1432 As, Norway
关键词
D O I
10.1074/jbc.273.52.35074
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Several series of site-directed mutations in thermolysin-like proteases are presented that show remarkable nonadditivity in their effect on thermal stability. A simple model is proposed that relates this nonadditivity to the occurrence of independent partial unfolding processes that occur in parallel at elevated temperatures. To prove this model, a thermolysin-like protease was designed in which two mutations located similar to 35 Angstrom apart in the structure individually exert small stabilizing effects of 2.3 and 4.1 degrees C, respectively, but when combined stabilize the protease by 14.6 degrees C. This overadditivity, which follows directly from the model, confirms that unfolding of this engineered protease starts in parallel at two different regions of the protein.
引用
收藏
页码:35074 / 35077
页数:4
相关论文
共 50 条
  • [31] Hydrolysis of industrial substrates by an extremely stable thermolysin-like protease variant obtained by protein engineering
    Van den Burg, B
    de Kreij, A
    Winkel, C
    Venema, G
    BIOTECHNOLOGY LETTERS, 1999, 21 (06) : 537 - 542
  • [32] THE UNFOLDING MECHANISM OF THERMOLYSIN
    CORBETT, RJT
    ROCHE, RS
    BIOPOLYMERS, 1983, 22 (01) : 101 - 105
  • [33] Purification, characterization, and primary structure of Clostridium perfringens lambda-toxin, a thermolysin-like metalloprotease
    Jin, F
    Matsushita, O
    Katayama, S
    Jin, SY
    Matsushita, C
    Minami, J
    Okabe, A
    INFECTION AND IMMUNITY, 1996, 64 (01) : 230 - 237
  • [34] THE ROLE OF HISTIDINE-231 IN THERMOLYSIN-LIKE ENZYMES - A SITE-DIRECTED MUTAGENESIS STUDY
    BEAUMONT, A
    ODONOHUE, MJ
    PAREDES, N
    ROUSSELET, N
    ASSICOT, M
    BOHUON, C
    FOURNIEZALUSKI, MC
    ROQUES, BP
    JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (28) : 16803 - 16808
  • [35] Hydrolysis of industrial substrates by an extremely stable thermolysin-like protease variant obtained by protein engineering
    B. Van den Burg
    A. de Kreij
    C. Winkel
    G. Venema
    Biotechnology Letters, 1999, 21 : 537 - 542
  • [36] THE ROLE OF THE PRO-SEQUENCE IN THE PROCESSING AND SECRETION OF THE THERMOLYSIN-LIKE NEUTRAL PROTEASE FROM BACILLUS-CEREUS
    WETMORE, DR
    WONG, SL
    ROCHE, RS
    MOLECULAR MICROBIOLOGY, 1992, 6 (12) : 1593 - 1604
  • [37] THERMOLYSIN-LIKE SERUM METALLOENDOPEPTIDASE - A NEW MARKER FOR ACTIVE SARCOIDOSIS THAT COMPLEMENTS SERUM ANGIOTENSIN-CONVERTING ENZYME
    ALMENOFF, J
    SKOVRON, ML
    TEIRSTEIN, AS
    ANNALS OF THE NEW YORK ACADEMY OF SCIENCES, 1986, 465 : 738 - 743
  • [38] Expression of Thermolysin-Like zinc metalloendopeptidases in vascular endothelial cells is regulated by shear stress and reactive oxygen species
    Fitzpatrick, PA
    Cotter, EJ
    Birney, YA
    Glucksman, MJ
    Cahill, PA
    Cummins, PM
    Franklin, R
    ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY, 2005, 25 (05) : E68 - E68
  • [39] CLINICAL-SIGNIFICANCE OF SERUM THERMOLYSIN-LIKE METALLOENDOPEPTIDASE AND ITS RELATIONSHIP TO SERUM ANGIOTENSIN CONVERTING ENZYME IN SARCOIDOSIS
    ALMENOFF, J
    TEIRSTEIN, AS
    AMERICAN JOURNAL OF MEDICINE, 1987, 82 (01): : 33 - 38
  • [40] DOMAIN UNFOLDING OF INTACT APO THERMOLYSIN
    CORBETT, RJT
    ROCHE, RS
    BIOPHYSICAL JOURNAL, 1985, 47 (02) : A181 - A181