Binding of nucleotide triphosphates to cardiotoxin analogue II from the Taiwan cobra venom (Naja naja atra). Elucidation of the structural interactions in the dATP-cardiotoxin analogue ii complex.
Snake venom cardiotoxins have been recently shown to block the enzymatic activity of phospholipid protein kinase and Na+,K+-ATPase. To understand the molecular basis for the inhibitory effects of cardiotoxin on the action of these enzymes, the nucleotide triphosphate binding ability of cardiotoxin analogue II (CTX II) from the Taiwan cobra (Naja naja atra) venom is investigated using a variety of spectroscopic techniques such as fluorescence, circular dichroism, and two-dimensional NMR. CTX II is found to bind to all the four nucleotide triphosphates (ATP, UTP, GTP, and CTP) with similar affinity. Detailed studies of the binding of dATP to CTX II indicated that the toxin molecule is significantly stabilized in the presence of the nucleotide. Molecular modeling, based on the NOEs observed for the dATP.CTX II complex, reveals that dATP binds to the CTX II molecule at the groove enclosed between the N- and C-terminal ends of the toxin molecule. Based on the results obtained in the present study, a molecular mechanism to account for the inhibition of the enzymatic activity of the phospholipid-sensitive protein kinase and Na+,K+-ATPase is also proposed.
Jayaraman
G
G
Department of Chemistry, National Tsing Hua University, Hsinchu, Taiwan, Republic of China.
Krishnaswamy
T
T
Kumar
S
S
Yu
C
C
D002942:Circular Dichroism; D004179:Cobra Cardiotoxin Proteins / Q000737:chemistry*; D003838:Deoxyadenine Nucleotides / Q000737:chemistry*; D004546:Elapid Venoms / Q000737:chemistry*; D005453:Fluorescence; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D009711:Nucleotides / Q000378:metabolism*; D011485:Protein Binding; D017510:Protein Folding; D047428:Protein Kinase Inhibitors; D017433:Protein Structure, Secondary; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors; D013696:Temperature; D014443:Tyrosine / Q000737:chemistry
D004179:Cobra Cardiotoxin Proteins; D003838:Deoxyadenine Nucleotides; D004546:Elapid Venoms; D009711:Nucleotides; D047428:Protein Kinase Inhibitors; D014443:Tyrosine; D000254:Sodium-Potassium-Exchanging ATPase; C026600:2'-deoxyadenosine triphosphate