Depth-dependent analysis of membranes using benzophenone-based phospholipids

  • John B
  • Kumar E
  • Lala A
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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.

Author-supplied keywords

  • Benzophenone-based phospholipids
  • Cross-linking
  • Depth-dependent analysis
  • Gaussian

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  • Bino John

  • E. Ravi Kumar

  • Anil K. Lala

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