Learning How to Make Bio-Active BAMLET
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Study Title:Effect of denaturation of alpha-lactalbumin on the formation of BAMLET (bovine alpha-lactalbumin made lethal to tumor cells).
A complex of alpha-lactalbumin with oleic acid, also known as HAMLET/BAMLET (human/bovine alpha-lactalbumin made lethal to tumor cells), causes apoptosis-like death in tumor cells but has little effect on healthy differentiated cells. The aim of this study was to examine whether irreversible denaturation of alpha-lactalbumin is detrimental to the formation and cytotoxicity of BAMLET. Commercial bovine holo alpha-lactalbumin (1-4% w/v) was heated at 80 degrees C for up to 100 min. With an increasing concentration of protein, the denaturation of alpha-lactalbumin proceeded faster, and aggregation became more extensive. Native and sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that a high proportion of the aggregates was linked by disulfide bonds. BAMLET was prepared from native and heat-treated alpha-lactalbumin according to a previously described chromatographic method. Despite the high content of denatured and aggregated alpha-lactalbumin in the heat-treated samples, their conversion into BAMLET was not negatively affected, resulting in BAMLET complexes partly composed of covalently linked aggregates of alpha-lactalbumin. The cytotoxicity of all prepared BAMLET samples was comparable to that of the control sample prepared from native alpha-lactalbumin (LD(50) = 34.6 +/- 2.7 mumol L(-1)). It was concluded that alpha-lactalbumin is not required to be in its native conformation for the conversion into its biologically active BAMLET complex.
Lisková K, Kelly AL, O'Brien N, Brodkorb A. Effect of denaturation of alpha-lactalbumin on the formation of BAMLET (bovine alpha-lactalbumin made lethal to tumor cells). J Agric Food Chem. 2010 April 58(7):4421-7.
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