title: Conformational and functional transitions in class II alpha-mannosidase from Aspergillus fischeri.
abstract: The conformational transitions in an oligomeric and high molecular weight class II alpha-mannosidase from Aspergillus fischeri were examined using fluorescence and CD spectroscopy under chemical, thermal and acid denaturing conditions. The enzyme lost the activity first and then the overall folded conformation and secondary structure. The midpoint values of GdnHCl mediated changes measured by inactivation; fluorescence and negative ellipticity were 0.48 M, 1.5 M and 1.9 M, respectively. The protein almost completely unfolded in 4.0 M GdnHCl but not at 90 degrees C. The inactivation and unfolding were irreversible. At pH 2.0, the protein exhibited molten-globule like intermediate with rearranged secondary and tertiary structures and exposed hydrophobic amino acids on the surface. This species showed increased accessibility of Trp to the quenchers and got denatured with GdnHCl in a different manner. The insoluble aggregates of a thermally denatured protein could be detected only in the presence of 0.25-0.75 M GdnHCl.
authors.affiliation: Biochemical Sciences Division, National Chemical Laboratory, Dr.Homi Bhabha road, Pune, 411008, India.
authors.forename: K S
authors.initials: KS
authors.lastname: Shashidhara
authors.forename: Sushama M
authors.initials: SM
authors.lastname: Gaikwad
mesh_terms: D001230:Aspergillus / Q000201:enzymology*; D002942:Circular Dichroism; D004789:Enzyme Activation; D019791:Guanidine / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008361:Mannosidases / Q000737:chemistry* / Q000378:metabolism*; D011489:Protein Denaturation / Q000187:drug effects; D017510:Protein Folding; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D013050:Spectrometry, Fluorescence; D013696:Temperature
chemical_list: D008361:Mannosidases; C038829:mannosyl-oligosaccharide 1,3 - 1,6-alpha-mannosidase; D019791:Guanidine