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Structural variations and molten globule state in Arisaema helliborifolium lectin under various treatments as monitored by spectroscopy

[ Vol. 23 , Issue. 2 ]


Kshema Thakur, Manpreet Kaur, Gulam Rabbani, Rizwan H. Khan, Sukhdev Singh and Jatinder Singh   Pages 107 - 119 ( 13 )


Solvent perturbation was used to study variations in structure of Arisaema helliborifolium lectin (AHL) with the help of circular dichroism (CD), intrinsic fluorescence (IF), extrinsic fluorescence, quenching and dynamic light scattering (DLS). AHL was studied under acidic, alkaline and 6 M guanidine hydrochloride (GuHCl) equilibrium states. Three structural states were identified for AHL at different conditions, that are native (N; pH 7.0), molten globule (MG; pH 2.0) and unfolded (U; pH 12.0). CD analysis revealed that 50% of secondary structure of AHL was -sheet component. A complete loss of secondary structure was observed at GuHCl treatment. The tertiary structural changes as studied by changes in microenvironment of trp residues also suggested a pH induced MG state as in case of CD. Parameter-A analysis pointed at the multi-step unfolding process of lectin under varying pH (pH 1-13). A comparision of CD and IF data further indicated that different pathways were followed for secondary and tertiary structure unfolding. Tryptophans of native AHL were only partially exposed to solvent belonging to Class II. Hydrodynamic diameter (Dh ) measurements of AHL via DLS also confirmed of a pH induced molten globule. A thermally induced molten globule was identified for AHL between 54-60 °C as monitored by DLS. An irreversible thermal denaturation was observed with the formation of a large aggregate. The Dh of AHL at neutral pH was confirmed by transmission electron microscopy (TEM).


Circular dichroism, Dynamic light scattering, Fluorescence, Guanidine hydrochloride, Lectin, Molten globule, pH denaturation, Thermal unfolding, Transmission electron microscopy.


Department of Molecular Biology and Biochemistry, Guru Nanak Dev University, Amritsar-143005, India

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