Expression and Biochemical Characterization of Acidic Phospholipase A(2)I from Agkistrodon acutus.
A cDNA encoding acidic phospholipase A(2)I(A.aAPLA(2)I)from Agkistrodon acutus was inserted into a bacterial expression vector and effectively expressed in E.coli RR1. The protein was produced as insoluble inclusion bodies. After partial purification by washing the inclusion bodies with Triton X-100, denaturing and refolding, the renatured recombinant protein was purified by FPLC column Superose(TM)12. The enzymatic acti-vity and platelet aggregation inhibiting effect of the expressed A.aAPLA(2)I is close to those of denatured-refolded native acidic PLA(2) from Agkistrodon halys Pallas, and has the same hemolytic activity as denatured-refolded basic phospholipase A(2) from Agkistrodon halys Pallas. The roles of various amino acid residues in the enzymatic activity and pharmacological activities of phospholipase A2 are discussed.
Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China. zhouyc@sunm.shcnc.ac.cn
Xiao-Long
XL
Liu
Xiang-Fu
XF
Wu
Ming-De
MD
Cai
Yuan-Cong
YC
Zhou