DIRECT EVALUATION OF STEREOSELECTIVITY OF CANCER ESTERASES BY POLYACRYLAMIDE-GEL ELECTROPHORESIS COUPLED WITH ACTIVITY STAINING WITH CHIRALNAPHTHYL ESTERS
Y. Yamazaki et al., DIRECT EVALUATION OF STEREOSELECTIVITY OF CANCER ESTERASES BY POLYACRYLAMIDE-GEL ELECTROPHORESIS COUPLED WITH ACTIVITY STAINING WITH CHIRALNAPHTHYL ESTERS, Analytical biochemistry, 231(2), 1995, pp. 295-300
Both enantiomers of alpha-naphthyl 2-phenylpropanoate (PhPr(ONap)), N-
acetylalaninate (AcAla(ONap)), N-methoxycarbonylalaninate (MocAla(ONap
)), N-methoxycarbonylvalinate (MocVal(ONap)), N-acetylprolinate (AcPro
(ONap)), and N-(trifluoroacetyl)prolinate (TfaPro(ONap)) were prepared
and used with Fast Blue RR salt for activity staining of esterases se
parated by polyacrylamide gel electrophoresis. Comparison of the band
thicknesses stained with each enantiomer indicates stereoselectivity o
f the major esterase in that band. Several esterases in normal rat liv
er, rat hepatoma-derived cells, and mouse B16 melanoma showed alterati
on or inversion of stereoselectivity by the substrate change from MocA
la(ONap) to MocVal(ONap) or from AcPro(ONap) to TfaPro(ONap). The ster
eoselectivity of the major esterases for MocVal(ONap) was reversed bet
ween the murine liver and the B16 melanoma enzymes. The present staini
ng with chiral naphthyl esters is very effective for surveying rapidly
the stereoselectivity of animal tissue and cancer esterases. (C) 1995
Academic Press, Inc.