We have previously introduced the use of permeabilized fission yeast cells(enzyme bags)that recombinantly express full-length CYPs for drug metabolism studies.Such enzyme bags are cells with pores that function as enz...We have previously introduced the use of permeabilized fission yeast cells(enzyme bags)that recombinantly express full-length CYPs for drug metabolism studies.Such enzyme bags are cells with pores that function as enzymes in situ.They can easily be prepared without a need for ultracentrifugation and may be used in similar protocols as microsomes.In this study we report the preparation of enzyme bag cocktails that permit the testing of multiple CYPs in a single enzyme bag reaction.Moreover,we established a convenient testing scheme that permits a rapid screen of all human CYPs for activity towards any given candidate substrate.An important aspect of this approach is the reduction of individual CYP test assays.If a cocktail containing many CYPs tests negative,it follows that all CYPs included in that cocktail need not be tested individually,thus saving time and resources.The new protocol was validated using two probe substrates.展开更多
Autoantibodies （aAbs） are generated by the immune system in response to a number of stimuli, one of which is believed to be aberrant protein targeting to the plasma surface. In this study, the presence of human cyto...Autoantibodies （aAbs） are generated by the immune system in response to a number of stimuli, one of which is believed to be aberrant protein targeting to the plasma surface. In this study, the presence of human cytochrome P450 enzyme CYP4Z1 on the outer surface of the plasma membrane of MCF-7 breast cancer cells is demonstrated by immunofluorescence. Moreover, anti- CYP4Z1 aAbs could clearly be detected in breast cancer patient sera by both immunoprecipitation and western blot analysis, while weak or no signals were obtained when testing controls.展开更多
文摘We have previously introduced the use of permeabilized fission yeast cells(enzyme bags)that recombinantly express full-length CYPs for drug metabolism studies.Such enzyme bags are cells with pores that function as enzymes in situ.They can easily be prepared without a need for ultracentrifugation and may be used in similar protocols as microsomes.In this study we report the preparation of enzyme bag cocktails that permit the testing of multiple CYPs in a single enzyme bag reaction.Moreover,we established a convenient testing scheme that permits a rapid screen of all human CYPs for activity towards any given candidate substrate.An important aspect of this approach is the reduction of individual CYP test assays.If a cocktail containing many CYPs tests negative,it follows that all CYPs included in that cocktail need not be tested individually,thus saving time and resources.The new protocol was validated using two probe substrates.
文摘Autoantibodies （aAbs） are generated by the immune system in response to a number of stimuli, one of which is believed to be aberrant protein targeting to the plasma surface. In this study, the presence of human cytochrome P450 enzyme CYP4Z1 on the outer surface of the plasma membrane of MCF-7 breast cancer cells is demonstrated by immunofluorescence. Moreover, anti- CYP4Z1 aAbs could clearly be detected in breast cancer patient sera by both immunoprecipitation and western blot analysis, while weak or no signals were obtained when testing controls.
