Structure of Ceramide-1-phosphate at the Air-water Solution Interface in the Absence and Presence of Ca2+Biophysical Journal
AbstractCeramide-1-phosphate, the phosphorylated form of ceramide, gained attention recently due to its diverse intracellular roles, in particular in inflammation mediated by cPLA2a. However, surprisingly little is known about the physical chemical properties of this lipid and its potential impact on physiological function. For example, the presence of Ca2þ is indispensable for the interaction of Cer-1-P with the C2 domain of cPLA2a. We report on the structure and morphology of Cer-1-P in monomolecular layers at the air/water solution interface in the absence and presence of Ca2þ using diverse biophysical techniques, including synchrotron x-ray reflectivity and grazing angle diffraction, to gain insight into the role and function of Cer-1-P in biomembranes. We show that relatively small changes in pH and the presence of monovalent cations dramatically affect the behavior of Cer-1-P. On pure water Cer-1-P forms a solid monolayer despite the negative charge of the phosphomonoester headgroup. In contrast, pH 7.2 buffer yields a considerably less solid-like monolayer, indicating that charge-charge repulsion becomes important at higher pH. Calcium was found to bind strongly to the headgroup of Cer-1-P even in the presence of a 100-fold larger Naþ concentration. Analysis of the x-ray reflectivity data allowed us to estimate how much Ca2þ is bound to the headgroup, ~0.5 Ca2þ and ~1.0 Ca2þ ions per Cer-1-P molecule for the water and buffer subphase respectively. These results can be qualitatively understood based on the molecular structure of Cer-1-P and the electrostatic/hydrogen-bond interactions of its phosphomonoester headgroup. Biological implications of our results are also discussed.
Citation InformationEdgar E Kooijman, David Vaknin, Wei Bu, Leela Joshi, et al.. "Structure of Ceramide-1-phosphate at the Air-water Solution Interface in the Absence and Presence of Ca2+" Biophysical Journal Vol. 96 Iss. 6 (2009) p. 2204 - 2215
Available at: http://works.bepress.com/wei_bu/2/