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Citation: LIU Shu-Qing, SUN Ming-Zhong, ZHAO Bao-Chang. Effects of Calcium Ion on Structural Integrity and Function of Phospholipase A₂[J]. Chinese Journal of Applied Chemistry, ;2009, 26(3): 257-262. shu

Effects of Calcium Ion on Structural Integrity and Function of Phospholipase A₂

1.
a Department of Biochemistry and Molecular Biology, Dalian Medical University College of Basic Medical Sciences, Dalian 116044;
2.
b Department of Biotechnology, Dalian Medical University College of Basic Medical Sciences, Dalian 116044

Corresponding author: SUN Ming-Zhong,
Received Date: 19 March 2008
Available Online: 8 May 2008
Fund Project: 国家自然科学基金(30512741) (30512741)大连医科大学高科技科研启动经费(20070301)资助项目 (20070301)

A novel phospholipase A₂(PLA₂) named GBB-PLA₂ was purified and determined by SDS-PAGE and mass spectrometry,from the snake venom of Glyodius blomhoffi Brevicaudaus.Electrophoresis,mass spectrometry,ICP-AES,UV-Vis and fluorescence spectroscopy techniques were utilized for the systematic study of GBB-PLA₂.The purified protein possessed molecular mass of 13 965.07 amu.Protein identification of GBB-PLA₂ was carried out by HPLC-ESI-MS/MS analysis.Four homogeneous peptides and 1 peptide of two PLA₂s Agkistrodon halys Pallas and Deinagkisrodon acutus snake venoms were identified from GBB-PLA₂.The results from native Sephacryl S-300 gel filtration and MALDI-TOF mass spectrometry indicate that GBB-PLA₂ exists as a stable monomeric molecule in aqueous solution.The experimental data from ICP-AES reveal that the purified GBB-PLA₂ is a Ca²⁺ bound protein with a calcium stoichiometry of 1:1([Ca²⁺]/[GBB-PLA₂]).Removal of Ca²⁺ from GBB-PLA₂ decreases the hydrolysis activity of PLA₂ by 90%,suggesting that Ca²⁺ ions play a critical role for GBB-PLA₂.The addition of exogenous Ca²⁺ can accelerate the hydrolysis rate of the substrate DPPC for GBB-PLA₂.Ca²⁺ increases the thermostability by 3 ℃ and changes the secondary structure of GBB-PLA₂,indicating that the presence of Ca²⁺ helps to maintain the structure integrity of GBB-PLA₂.
- phospholipase A₂,
- calcium ion,
- structure,
- function
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Effects of Calcium Ion on Structural Integrity and Function of Phospholipase A₂