title: Study of interaction of GM2 activator protein with GM2 using circular dichroism and fluorescence spectroscopy.
abstract: GM2 activator protein (GM2AP) is a cofactor for stimulating the enzymatic hydrolysis of the glycolipid GM2 by beta-hexosaminidase A to produce GM3. We have examined the conformation of GM2AP before and after its interaction with GM2, GM3, and GA2 using circular dichroism and fluorescence spectroscopy techniques. In the presence of GM2, a blue shift of the fluorescence emission maximum and a strong decrease of molar ellipticity values in circular dichroism spectra were observed only at pH 4.5 and at GM2/GM2AP molar ratio higher than 10:1 (up to 50:1). These results suggest that GM2AP assumed a more organized alpha-helical conformation with the tryptophan residues moving from the polar medium toward the hydrophobic environment of the protein. The conformation of GM2AP in the presence of the downstream reaction product, GM3, or a less favorable substrate, GA2, clearly differed from that in the presence of GM2. The relationships between spectroscopic changes and enzymatic activity, herein discussed, strongly suggest that the specific conformation exhibited by GM2AP in the presence of GM2 is functional to serve as an activator for the enzymatic hydrolysis of GM2.
authors.affiliation: Department of Experimental, Environmental Medicine and Biotechnology, University of Milano-Bicocca, Monza, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Ravasi
authors.forename: Massimo
authors.initials: M
authors.lastname: Masserini
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Vecchio
authors.forename: Yu-Teh
authors.initials: YT
authors.lastname: Li
authors.forename: Su-Chen
authors.initials: SC
authors.lastname: Li
mesh_terms: D002942:Circular Dichroism; D049289:G(M2) Activator Protein; D005678:G(M2) Ganglioside / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
chemical_list: D049289:G(M2) Activator Protein; D011506:Proteins; D005678:G(M2) Ganglioside