[The hemolytic site of the basic phospholipase A(2) from Agkistrodon halys pallas].
The gene of the basic phospholipase A(2) from Agkistrodon halys Pallas (BPLA(2) )was mutated site-directedly by polymerase chain reaction (PCR) and the residue Arg(34) of the encloding protein was substituted by Glu and Gln   respectively. The mutant gene has been cloned into the expression vector pBLMVL2 and has been expressed in E.coli RR1 effectively. The protein was produced as insoluble inclusion bodies. After partial purification, the inclusion bodies were denatured and renatured into active form, and the renatured recombinant protein was purified by gel-filtration. The expression product has the same enzymatic activity as the denatured-refolded BPLA(2) and its hemolytic activity dropped distinctly, which suggest that the basic residue Arg(34) of BPLA(2) is a crucial amino acid residue during the process of hemolysis.
Jiao
H M
HM
Institute of Biochemistry and Cell Biology, the Chinese Academy of Sciences, Shanghai 200031, China.
Jin
Q
Q
Zhao
J J
JJ
Feng
B
B
Wu
X F
XF
Zhou
Y C
YC
D017836:Agkistrodon; D019943:Amino Acid Substitution; D000818:Animals; D001120:Arginine / Q000235:genetics / Q000378:metabolism; D006461:Hemolysis / Q000502:physiology*; D016297:Mutagenesis, Site-Directed; D010741:Phospholipases A / Q000235:genetics / Q000378:metabolism*
D001120:Arginine; D010741:Phospholipases A