Expression, purification and functional characterization of a recombinant scorpion venom peptide BmTXKbeta.
BmTXKbeta, a scorpion toxin isolated from the Chinese scorpion Buthus martensii Karsch (BmK), was expressed as a GST fusion protein in BL21 (DE3) strain. The recombinant GST-BmTXKbeta protein was purified by affinity chromatography. When treated with enterokinase, the GST-BmTXKbeta fusion protein released an approximate 6.5kDa protein which was the expected size for correctly processed. About 2mg purified recombinant BmTXKbeta protein (rBmTXKbeta) was produced from 1l bacterial culture, using this expression and purification system. The function of rBmTXKbeta was studied on the rabbit atrial myocyte by whole-cell patch clamp technique. The results showed that rBmTXKbeta inhibited the transient outward current (I(to)) of rabbit atrial myocyte with recovery after washout and the inhibition was concentration-dependent. The rBmTXKbeta prolonged the action potential duration of rabbit atrial myocyte in a concentration-dependent manner, whereas it did not affect the action potential amplitude.
Cao
Zhijian
Z
Department of Biotechnology, Institute of Virology, College of Life Science, Wuhan University, 430072, Wuhan, PR China.
Xiao
Fan
F
Peng
Fang
F
Jiang
Dahe
D
Mao
Xin
X
Liu
Hui
H
Li
Wenxin
W
Hu
Dan
D
Wang
Teng
T
D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000502:physiology; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D012604:Scorpion Venoms / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology
C121628:BmTXKbeta1 protein, Buthus martensii; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D012604:Scorpion Venoms