Functions, structures and Triton X-100 effect for the catalytic subunits of heterodimeric phospholipases A2 from Vipera nikolskii venom.
Phospholipases A(2) (PLA(2)s) from snake venoms have diverse pharmacological functions including neurotoxicity, and more studies are necessary to understand relevant mechanisms. Here we report the different crystal structures for two enzymatically active basic subunits (HDP-1P and HDP-2P) of heterodimeric neurotoxic PLA(2)s isolated from Vipera nikolskii venom. Structural comparisons with similar PLA(2)s clearly show some flexible regions which might be important for the catalytic function and neurotoxicity. Unexpectedly, Triton X-100 molecule bound in the hydrophobic channel of HDP-1P and HDP-2P was observed, and its binding induced conformational changes in the Ca(2+) binding loop. Enzymatic activity measurements indicated that Triton X-100 decreased the activity of PLA(2), although with comparatively low inhibitory activity. For the first time exocytosis experiments in pancreatic beta cells were used to confirm the presynaptic neurotoxicity of relevant snake PLA(2). These experiments also indicated that Triton X-100 inhibited the influence of HDP-1P on exocytosis, but the inhibition was smaller than that of MJ33, a phospholipid-analogue inhibitor of PLA(2). Our studies performed at a cellular level are in good agreement with earlier findings that enzymatic activity of the snake presynaptic PLA(2) neurotoxins is essential for effective block of nerve terminals.
Gao
Wei
W
Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101, China.
Starkov
Vladislav G
VG
He
Zi-Xuan
ZX
Wang
Qi-hai
QH
Tsetlin
Victor I
VI
Utkin
Yuri N
YN
Lin
Zheng-jiong
ZJ
Bi
Ru-chang
RC
D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D002460:Cell Line; D003460:Crystallization; D019281:Dimerization; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017830:Octoxynol / Q000737:chemistry*; D054467:Phospholipases A2 / Q000737:chemistry* / Q000302:isolation & purification / Q000633:toxicity*; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D014757:Viper Venoms / Q000201:enzymology*
D014757:Viper Venoms; D017830:Octoxynol; D054467:Phospholipases A2