Open AccessCleavage of the synaptobrevin/vesicle‐associated membrane protein (VAMP) of the mouse brain by the recombinant light chain of Clostridium botulinum type B toxin
Author(s) -
Rhee Sang Dal,
Jung Hyun Ho,
Yang GiHyeok,
Moon Yu Seok,
Yang KyuHwan
Publication year - 1997
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1997.tb10371.x
Subject(s) - synaptobrevin , recombinant dna , cleavage (geology) , toxin , clostridium botulinum , immunoglobulin light chain , chemistry , microbiology and biotechnology , biology , vesicle , synaptic vesicle , biochemistry , membrane , gene , genetics , antibody , paleontology , fracture (geology)
The light chain of Clostridium botulinum type B toxin was expressed in Escherichia coli using the expression vector pET‐3a containing phage T 7 promoter. The expressed protein was then purified by DEAE‐cellulose and phosphocellulose chromatography and the proteolytic activity of the purified light chain was studied. The purified recombinant light chain cleaved synaptobrevin when mixed with the mouse brain microsome and the proteolytic activity of the light chain was inhibited if a metal chelating agent such as EDTA or 2,2′‐dipyridyl was added. The recombinant light chain cleaved synaptobrevin more effectively than the native type B toxin. When the native toxin was trypsinized and was reduced with DTT, its proteolytic activity was similar to that of the recombinant light chain.