Peripheral binding mode and penetration depth of cobra cardiotoxin on phospholipid membranes as studied by a combined FTIR and computer simulation approach.
Cobra cardiotoxin, a cytotoxic beta-sheet basic polypeptide, is known to cause membrane leakage in many cells including human erythrocytes. Herein, we demonstrate that the major cobra cardiotoxin from Naja atra, CTX A3, can cause leakage of vesicle contents in phosphatidylglycerol (PG) and phosphatidylserine containing, but not in pure phosphatidylcholine (PC), membrane bilayers. By the combined polarized attenuated total reflection infrared spectroscopy and computer simulation studies, CTX A3 is shown to peripherally bind to both zwitterionic and anionic monolayers in a similar edgewise manner with a tilted angle of approximately 48 +/- 20 degrees between the beta-sheet plane of the CTX molecule and the normal of the membrane surface. The average surface area expansion induced by CTX A3 binding to the PG monolayer, however, is two times larger than that of the PC monolayer as determined by the Langmuir minitrough method. Interaction energy considerations of CTX A3 on neutral and negatively charged membrane surfaces suggests that the electrostatic interaction between anionic lipid and cationic CTXs plays a role in modulating the penetration depth of CTX molecules on the initial peripheral binding mode and reveals a pathway leading to the formation of an inserted mode in negatively charged membrane bilayers.
Huang
Wei-Ning
WN
Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu, 30043, Taiwan.
Sue
Shih-Che
SC
Wang
Da-Shin
DS
Wu
Po-Long
PL
Wu
Wen-guey
WG
D000818:Animals; D004179:Cobra Cardiotoxin Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D003198:Computer Simulation; D020404:Glycerophospholipids / Q000737:chemistry* / Q000378:metabolism*; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism*; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism*; D008564:Membrane Potentials; D008567:Membranes, Artificial; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D011312:Pressure; D011485:Protein Binding / Q000187:drug effects; D017433:Protein Structure, Secondary; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods; D055672:Static Electricity; D013816:Thermodynamics
D004179:Cobra Cardiotoxin Proteins; D020404:Glycerophospholipids; D008051:Lipid Bilayers; D008563:Membrane Lipids; D008567:Membranes, Artificial