Depth-dependent analysis of membranes using benzophenone-based phospholipids

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Abstract

Any attempt to probe the membrane hydrophobic core with chemical reagents necessitates the use of reactive intermediates like carbenes and nitrenes, which can insert into C-H bonds. Several photoactivable reagents based on carbenes and nitrenes have been reported. However, the high reactivity of these reagents, often leads to very low insertion yields. We report here a high degree of cross-linking (35-40%) achieved with three benzophenone-based phospholipids and analyze the carbon functionalization data using a multiple Gaussian function. These phospholipids are so designed so as to permit depth-dependent labeling in membranes. Single bilayer vesicles were prepared from these phospholipids and dimyristoylphosphatidylcholine. The cross-linked product was isolated and characterized by mass spectroscopy. The results obtained indicated that the cross-linked product was dominated by dimeric product formed by intermolecular cross-linking. The Gaussian analysis used here provides insight into the relative depths of the probes inside the membrane. (C) 2000 Elsevier Science B.V.

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John, B., Kumar, E. R., & Lala, A. K. (2000). Depth-dependent analysis of membranes using benzophenone-based phospholipids. Biophysical Chemistry, 87(1), 37–42. https://doi.org/10.1016/S0301-4622(00)00178-2

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