@article { ISI:000250755100004,
	title = {Fluorescence quenching and time-resolved fluorescence studies of alpha-Mannosidase from Aspergillus fischeri (NCIM 508)},
	journal = {Journal of Fluorescence},
	volume = {17},
	number = {6},
	year = {2007},
	month = {NOV},
	pages = {599-605},
	publisher = {SPRINGER/PLENUM PUBLISHERS},
	type = {Article},
	address = {233 SPRING ST, NEW YORK, NY 10013 USA},
	abstract = {<p>Apart from the vital role in glycoprotein biosynthesis and degradation, alpha-mannosidase is currently an important therapeutic target for the development of anticancer agents. Fluorescence quenching and time-resolved fluorescence of alpha-mannosidase, a multitryptophan protein from Aspergillus fischeri were carried out to investigate the tryptophan environment. The tryptophans were found to be differentially exposed to the solvent and were not fully accessible to the neutral quencher indicating heterogeneity in the environment. Quenching of the fluorescence by acrylamide was collisional. Surface tryptophans were found to have predominantly positively charged amino acids around them and differentially accessible to the ionic quenchers. Denaturation led to more exposure of tryptophans to the solvent and consequently in the significant increase in quenching with all the quenchers. The native enzyme showed two different lifetimes, tau (1) (1.51 ns) and tau (2) (5.99 ns). The average lifetime of the native protein (tau) (3.187 ns) was not affected much after denaturation (tau) (3.219 ns), while average lifetime of the quenched protein samples was drastically reduced (1.995 ns for acrylamide and 1.537 ns for iodide). This is an attempt towards the conformational studies of alpha-mannosidase.</p>},
	keywords = {alpha-Mannosidase, Denaturation, fluorescence, lifetime, Solute quenching},
	issn = {1053-0509},
	doi = {10.1007/s10895-007-0227-8},
	author = {Shashidhara, K. S. and Gaikwad, Sushama M.}
}
@article { ISI:000280597500004,
	title = {Conformational and functional transitions in class II alpha-mannosidase from aspergillus fischeri},
	journal = {Journal of Fluorescence},
	volume = {20},
	number = {4},
	year = {2010},
	month = {JUL},
	pages = {827-836},
	publisher = {SPRINGER/PLENUM PUBLISHERS},
	address = {233 SPRING ST, NEW YORK, NY 10013 USA},
	abstract = {<p>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 A 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.</p>},
	keywords = {Aggregation, alpha-Mannosidase, ANS binding, Circular dichroism, fluorescence, GdnHCl, Hydrobicity, Thermal denaturation},
	issn = {1053-0509},
	doi = {10.1007/s10895-010-0625-1},
	author = {Shashidhara, K. S. and Gaikwad, Sushama M.}
}
