31P NMR and AFM studies on the destabilization of cell and model membranes by the major bovine seminal plasma protein, PDC-109.
The effect of PDC-109 binding to dimyristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylglycerol (DPPG) multilamellar vesicles (MLVs) and supported membranes was investigated by (31)P NMR spectroscopy and atomic force microscopy. Additionally, the effect of cholesterol on the binding of PDC-109 to phosphatidylcholine (PC) membranes was studied. Binding of PDC-109 to MLVs of DMPC and DPPG induced the formation of an isotropic signal in their (31)P NMR spectra, which increased with increasing protein/lipid ratio and temperature, consistent with protein-induced disruption of the MLVs and the formation of small unilamellar vesicles or micelles but not inverse hexagonal or cubic phases. Incorporation of cholesterol in the DMPC MLVs afforded a partial stabilization of the lamellar structure, consistent with previous reports of membrane stabilization by cholesterol. AFM results are consistent with the above findings and show that addition of PDC-109 leads to a complete breakdown of PC membranes. The fraction of isotropic signal in (31)P NMR spectra of DPPG in the presence of PDC-109 was less than that of DMPC under similar conditions, suggesting a significantly higher affinity of the protein for PC. Confocal microscopic studies showed that addition of PDC-109 to human erythrocytes results in a disruption of the plasma membrane and release of hemoglobin into the solution, which was dependent on the protein concentration and incubation time.
School of Chemistry, University of Hyderabad, Hyderabad - 500 046, India.
Rajani S
RS
Damai
Rajeshwer S
RS
Sankhala
Veerappan
V
Anbazhagan
Musti J
MJ
Swamy
D000818:Animals; D016880:Anisotropy; D002417:Cattle; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D004910:Erythrocyte Membrane / Q000187:drug effects*; D006801:Humans; D008081:Liposomes / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D018625:Microscopy, Atomic Force; D018613:Microscopy, Confocal; D010715:Phosphatidylglycerols / Q000737:chemistry; D010761:Phosphorus Radioisotopes; D029607:Seminal Vesicle Secretory Proteins / Q000494:pharmacology*
D008081:Liposomes; D010715:Phosphatidylglycerols; D010761:Phosphorus Radioisotopes; D029607:Seminal Vesicle Secretory Proteins; C424303:seminal vesicle secretory protein 109, Bos taurus; D004134:Dimyristoylphosphatidylcholine; C030345:1,2-dipalmitoylphosphatidylglycerol