Biophysical characterization of the stability of the 150-kilodalton botulinum toxin, the nontoxic component, and the 900-kilodalton botulinum toxin complex species

被引:75
作者
Chen, F
Kuziemko, GM
Stevens, RC [1 ]
机构
[1] Univ Calif Berkeley, Dept Chem, Berkeley, CA 94720 USA
[2] Univ Calif Berkeley, Grad Grp Biophys, Berkeley, CA 94720 USA
[3] Univ Calif Berkeley, Lawrence Berkeley Lab, Div Mat Sci, Berkeley, CA 94720 USA
关键词
D O I
10.1128/IAI.66.6.2420-2425.1998
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Botulinum neurotoxin serotype A is initially released from the bacterium Clostridium botulinum as a stable 900-kDa complex. The serotype A 900-kDa complex is one of the forms of the toxin being used as a therapeutic agent for the treatment of various neuromuscular disorders. Previous experiments have demonstrated that the 900-kDa complex form of the toxin protects the toxin from the harsh conditions of the gastrointestinal tract. To provide molecular level details of the stability and equilibrium of the 900-kDa complex, the nontoxic component, and the toxic (botulinum neurotoxin) component, the three species have been investigated,vith a series of biophysical techniques at the molecular level (dynamic light scattering, proteolysis, circular dichroism, pH incubations, and agglutination assays). These experiments were conducted under harsh conditions which mimic those found along the gastrointestinal tract. Separately, exposure to denaturing and proteolytic conditions degrades both the botulinum neurotoxin and the nontoxic component. In the 900-kDa complex, the botulinum neurotoxin is protected during exposure to the gastrointestinal environment and the nontoxic component is slightly modified. Surprisingly, the toxin protects the ability of the nontoxic component to agglutinate erythrocytes. Contrary to previous reports, the purified 900-kDa complex did not have agglutination ability until after exposure to the proteolytic conditions. These experiments provide new evidence and detail for the theory that the nontoxic component and the toxic component protect one another during exposure to harsh conditions, and a molecular model is presented for the passage of the toxin through the gastrointestinal tract.
引用
收藏
页码:2420 / 2425
页数:6
相关论文
共 36 条
  • [1] BALDING P, 1973, IMMUNOLOGY, V25, P773
  • [2] BECK WS, 1991, LIFE INTRO BIOL, P666
  • [3] BOTULINUM NEUROTOXIN-A SELECTIVELY CLEAVES THE SYNAPTIC PROTEIN SNAP-25
    BLASI, J
    CHAPMAN, ER
    LINK, E
    BINZ, T
    YAMASAKI, S
    DECAMILLI, P
    SUDHOF, TC
    NIEMANN, H
    JAHN, R
    [J]. NATURE, 1993, 365 (6442) : 160 - 163
  • [4] THE N-TERMINAL HALF OF THE HEAVY-CHAIN OF BOTULINUM TYPE A NEUROTOXIN FORMS CHANNELS IN PLANAR PHOSPHOLIPID-BILAYERS
    BLAUSTEIN, RO
    GERMANN, WJ
    FINKELSTEIN, A
    DASGUPTA, BR
    [J]. FEBS LETTERS, 1987, 226 (01) : 115 - 120
  • [5] Antibody mapping to domains of botulinum neurotoxin serotype A in the complexed and uncomplexed forms
    Chen, F
    Kuziemko, GM
    Amersdorfer, P
    Wong, C
    Marks, JD
    Stevens, RC
    [J]. INFECTION AND IMMUNITY, 1997, 65 (05) : 1626 - 1630
  • [6] CHROMATOGRAPHIC FRACTIONATION OF CRYSTALLINE TOXIN CLOSTRIDIUM BOTULINUM TYPE A
    DASGUPTA, BR
    BOROFF, DA
    ROTHSTEI.E
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1966, 22 (06) : 750 - &
  • [7] DASGUPTA BR, 1968, J BIOL CHEM, V243, P1065
  • [8] CIRCULAR DICHROIC AND FLUORESCENCE SPECTROSCOPIC STUDY OF THE CONFORMATION OF BOTULINUM NEUROTOXIN TYPE-A AND TYPE-E
    DATTA, A
    DASGUPTA, BR
    [J]. MOLECULAR AND CELLULAR BIOCHEMISTRY, 1988, 79 (02) : 153 - 159
  • [9] PROTEIN-LIPID INTERACTIONS IN HUMAN SMALL INTESTINAL BRUSH-BORDER MEMBRANES
    DUDEJA, PK
    HARIG, JM
    RAMASWAMY, K
    BRASITUS, TA
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY, 1989, 257 (05): : G809 - G817
  • [10] PEPTIDE AND PROTEIN MOLECULAR-WEIGHT DETERMINATION BY ELECTROPHORESIS USING A HIGH-MOLARITY TRIS BUFFER SYSTEM WITHOUT UREA
    FLING, SP
    GREGERSON, DS
    [J]. ANALYTICAL BIOCHEMISTRY, 1986, 155 (01) : 83 - 88