Effect of chaotropic agents on the structure-function of recombinant acylpeptide hydrolase.
作者： R. Senthilkumar , K. Krishna Sharma
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摘要: Acylpeptide hydrolase, a new class the serine-type peptidase, belongs to alpha,beta hydrolase group of proteins. The tetrameric enzyme showed varying degree stability in presence 1-8 M urea. displayed about 15% its original activity when treated with 8 urea for 1 h at 25 degrees C. Complete recovery was observed on dialysis or dilution (50-fold) denatured enzyme. However, complete abolition GnHCl. Dialysis GnHCl-treated resulted 15-20% activity. fluorescence emission spectra native 337 nm red shift up 16 and 18 4 Native during far-UV circular dichroism spectroscopy exhibited predominantly beta-sheet structure. lost secondary structure concentrations 2 higher, whereas tertiary minimally perturbed below GnHCl both structures found dissociate into monomers 70 kDa. Both monomeric dimeric species were after 24-h indicating reassociation process. monomer dimers forms recovered active.
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