Characterization of complex formation between lipopolysaccharide and lysozyme.
The binding of lysozyme (LZM) to bacterial lipopolysaccharide (LPS) inhibited the biological activities of LPS as well as the enzymic activity of LZM. The mode of binding has been characterized by using dansylated LZM and enzyme inhibition. The binding of LPS to LZM significantly increased the fluorescence intensity (Fl-intensity) of the danyl group and was found to be time-dependent; the complex was produced gradually and became stabilized within 20 min at 37 degrees, 10 min at 50 degrees, and 1 min at 70 degrees. The maximum level of binding was also dependent on the reaction temperature, and more complex was formed at higher temperatures. Complexation was strongly dependent on the salt concentration and was not observed at greater than 0.5M NaCl. From collected evidence of the Fl-intensities of various dansyl derivatives and amphiphiles, it is concluded that LZM interacts with LPS by multiple binding-modes, the first being strongly related to the enzyme inhibition, the second being close to the Fl-intensity, and the third being dependent on the inhibition of immunopharmacological activities. For the amphiphiles used in this study, sodium dodecyl sulfate (SDS), 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), 3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-propanesulfonate (CHAPSO), decansulfonic acid, and cardiolipin have binding modes similar to that of LPS.
Ohno
N
N
Laboratory for Immunopharmacology of Microbial Products, Tokyo College of Pharmacy, Japan.
Tanida
N
N
Yadomae
T
T
D000345:Affinity Labels; D000818:Animals; D003619:Dansyl Compounds; D003902:Detergents / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007700:Kinetics; D008070:Lipopolysaccharides / Q000378:metabolism*; D009113:Muramidase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009994:Osmolar Concentration; D013050:Spectrometry, Fluorescence; D013696:Temperature
D000345:Affinity Labels; D003619:Dansyl Compounds; D003902:Detergents; D008070:Lipopolysaccharides; D009113:Muramidase; C029038:dansyl chloride