Glycogen β-particles surface characterized by a combination of size exclusion chromatography and pyrene excimer fluorescence before and after β-amylolysis

Abstract

Size exclusion chromatography (SEC) and pyrene excimer formation (PEF) experiments were conducted to characterize the local density profile inside a glycogen sample before (Glycogen) and after (Gly-β-LD) treatment with β-amylase. These experiments were conducted to assess whether the density at the periphery of the glycogen particles was very high to limit access to proteins involved in the metabolism of glycogen as predicted by the Tier model or low as suggested by the Gilbert model. SEC analysis indicated that the density inside the Glycogen and Gly-β-LD samples remained constant with particle size and was not affected by β-amylolysis. Analysis of the PEF experiments conducted on the Glycogen and Gly-β-LD samples labeled with 1-pyrenebutyric acid showed that the particles have a dense interior and loose corona. The conclusions reached by the SEC and PEF experiments agree with the Gilbert model and have implications for the association of glycogen β-particles into larger α-particles.

Keywords: Glycogen; Polysaccharides; Pyrene excimer formation; β-Amylolysis.