Changes in the Fluorescence Characteristics of N-(1-Pyrene) Maleimide Bound to the Intestinal Brush-Border Membranes by Neuraminidase Treatment

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The effects of neuraminidase treatment on the dynamic properties of the porcine intestinal brush-border membranes have been examined by using a fluorogenic thiol reagent, N-(1-pyrene)maleimide (NPM). Desialylation of the membranes by treatment with neuraminidase resulted in changes in the fluorescence parameters of NPM-labeled membranes, i.e. a decrease of the fluorescence lifetime and a suppression of the temperature-dependent decrease of the fluorescence intensity. These results suggest that the environmental properties around NPM-labeled SH groups in the membrane proteins were modified by neuraminidase treatment. Perturbation of the microenvironment around NPM-labeled SH groups associated with desialylation by the enzyme treatment was also determined by measuring the increase of fluorescence anisotropy and decrease of quenching efficiency with acrylamide or CH_3COOTl of the complex. Based on the results it is suggested that the dynamic properties of the conformation around NPM-labeled SH groups in the membrane proteins are sensitively influenced by neuraminidase treatment.
社団法人日本薬学会の論文
1989-08-25