Antisera probes to an atypical pseudocholinesterase from surgeonfish reveal immunochemical variability and tissue-specific molecular polymorphism.
Polyclonal antisera were raised in rabbits against the purified sialated, presumed-globular tetrameric pseudocholinesterase (pseudo-ChE) from surgeonfish (Leibel: Journal of Experimental Zoology 1988b) and against commercially obtained Electrophorus electroplax AChE. The resulting antisera probes were absolutely specific for their respective antigens and failed to titrate ChE activities heterologously. However, each antisera probe did crossreact with its other respective globular and asymmetric aggregational isozymes. The resultant specific probes were then used to examine interspecific evolutionary conservation of the two ChE activities and, in conjunction with velocity sedimentation analysis and differential paraoxon inhibition, the tissue distribution and molecular polymorphism of these same two enzyme systems in surgeonfish. These experiments suggest the tight evolutionary conservation of AChE in contrast to the apparent high variability of pseudo-ChE amongst the wide range of teleost fishes tested. The native atypical pseudo-ChE was shown to exist, like AChE, as a series of sialated and asialated globular and asymmetric aggregational isozymes whose relative distribution exhibits marked tissue specificity. The extremely high levels of pseudo-ChE characteristic of white skeletal (epaxial) muscle, in particular, was conspicuous, and its occurrence in the sarcolemma is discussed in the context of its possible function and in relation to the apparent lack of evolutionary conservation amongst marine teleosts.
Leibel
W S
WS
Department of Biology, Lafayette College, Easton, Pennsylvania 18042.
D000818:Animals; D000918:Antibody Specificity; D000940:Antigenic Variation; D005075:Biological Evolution; D001921:Brain / Q000201:enzymology; D002091:Butyrylcholinesterase / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D002499:Centrifugation, Density Gradient; D002802:Cholinesterases / Q000032:analysis*; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D005123:Eye / Q000201:enzymology; D005399:Fishes / Q000378:metabolism*; D007106:Immune Sera / Q000276:immunology; D007122:Immunoelectrophoresis; D007150:Immunohistochemistry; D008099:Liver / Q000201:enzymology; D009132:Muscles / Q000201:enzymology; D011110:Polymorphism, Genetic
D007106:Immune Sera; D002091:Butyrylcholinesterase; D002802:Cholinesterases