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Resistance of Bromelain to SDS Binding

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Title Resistance of Bromelain to SDS Binding
 
Creator Bhattacharya, Reema
Bhattacharyya, Debasish
 
Subject Structural Biology & Bioinformatics
 
Description Interaction of the plant cysteine protease bromelain with SDS has been studied using CD spectroscopy,
intrinsic fluorescence emission, extrinsic fluorescence probe pyrene, isothermal calorimetric (ITC)
investigations and inhibition of hydrolyzing activity. Results exhibit number of synchronous transitions
when plotted against the total SDS concentration. SDS at submicellar level caused conformation change of
bromelain leading to a stable entity. ITC and pyrene experiments suggest that the structural modifications
below 5 mM, the cmcapp of SDS solutions containing bromelain, are the result of alterations of solvent
hydrophobicity or non-specific weak binding and/or adsorption of SDS monomers. Melting temperature
(Tm) and the free energy change for thermal unfolding (ΔGunf) of the SDS induced conformers was
decreased by 5 °C and 0.5 kcal/mol respectively, compared to native bromelain. Below 5 mM, SDS caused
large decrease in Vmax without affecting Km for the substrate Z-Arg-Arg-NHMec. Analysis of kinetic data
imply that SDS acts as a partial non-competitive inhibitor since even at 100 mM, the residual activity of
bromelain was retained by 3%. Inhibition studies show an IC50 of 0.55 mM and a high Ki of 0.145 mM.
These demonstrate that bromelain is resistant to SDS binding and denaturation, a property known for β-
sheet rich kinetically stable proteins
 
Publisher Elsevier
 
Date 2009
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/1358/1/BIOCHIMICA_ET_BIOPHYSICA_ACTA%2DPROTEINS_AND_PROTEOMICS__1794(_4)_698%2D708;2009[88].pdf
Bhattacharya, Reema and Bhattacharyya, Debasish (2009) Resistance of Bromelain to SDS Binding. BBA - Biochimica et Biophysica Acta, 1794 (4). pp. 698-708.
 
Relation http://dx.doi.org/10.1016/j.bbapap.2008.12.019
http://www.eprints.iicb.res.in/1358/