New synthesis and conformational analysis of lipid A: biological activity and supramolecular assembly

Abstract

In order to investigate precise structural requirements for expression of the biological activity of lipid A, new structural analogs were synthesized via improved, highly efficient preparative routes. The number and chain length of the fatty acyl groups proved to have crucial influence on the biological activity. For the study of conformation relevant to activity of lipid A by means of NMR spectrometry, lipid A analogs [13C]-labeled at the 6-position were then synthesized starting from 6-[13C]-glucose. Precise analysis of J CH and NOE data disclosed a particular molecular conformation and characteristic supramolecular assembly in an aqueous SDS micelle solution of the tetraacyl biosynthetic precursor molecule. The observed mode of assembly may reflect the actual behavior of lipid A molecules in the cell surface outer membrane of living bacteria. A biologically inactive, artificial, tetra-acyl analog having shorter acyl chains exhibits, by contrast, distinct conformations with no sign of supramolecule formation. The acyl moieties in lipid A are thus found to play an important role in regulating the overall conformation of the hydrophilic backbone.