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Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy
Published on: August 20, 2018
Hydrophobic interactions are the prevalent force in bromelain:Fab' complex
P Gupta1, M Saleemuddin, R H Khan
1Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, Uttar Pradesh 202002, India. pawan_g75@hotmail.com
Abstract:
Antibromelain polyclonal antibodies against stem bromelain were raised in male albino rabbits and the Fab monomers isolated from the IgG of the immune sera as reported in our earlier communication (Gupta, P., Khan, R. H., and Saleemuddin, M. (2003) Biochim. Biophys. Acta, 1646, 131-135). Further, as evident from that communication bromelain:Fab complex has 1 : 1 stoichiometry. The stability of bromelain:Fab complex (1 : 1 stoichiometry) was investigated by far and near-UV CD and fluorescence measurements. Addition of up to 1.8 M NaCl caused no significant changes in fluorescence signals and near-UV CD peak pattern. However, the spectral studies together with gel filtration studies suggest dissociation of the complex beyond 5% (v/v) methanol. These results show that hydrophobic interactions play a pronounced role in the binding of Fab to bromelain while electrostatic interactions may be less crucial.
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