. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. V-ATPASE IN THE ACCESSORY BORING ORGAN OF A MURIC1D GASTROPOD 281 name "single celled epithelia" has been coined (Brown and Breton, 1996). SDS-PAGE and immunoblotting provided additional im- munological evidence to support the identity of V-ATPase pumps. Gel electrophoresis revealed that a membrane prep- aration of ABO homogenate contains peptides with molec- ular weights that are similar to those of V-ATPase subunits. Additional bands in the gel reflect the impurity of our sample. Furthermore, dot and western immu
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. V-ATPASE IN THE ACCESSORY BORING ORGAN OF A MURIC1D GASTROPOD 281 name "single celled epithelia" has been coined (Brown and Breton, 1996). SDS-PAGE and immunoblotting provided additional im- munological evidence to support the identity of V-ATPase pumps. Gel electrophoresis revealed that a membrane prep- aration of ABO homogenate contains peptides with molec- ular weights that are similar to those of V-ATPase subunits. Additional bands in the gel reflect the impurity of our sample. Furthermore, dot and western immunoblots of sam- ples derived from ABO homogenates stained positive for V-ATPase. The western blots show that polyclonal antibod- ies raised against mammalian V-ATPase subunits cross- react to the equivalent peptides of V-ATPases found in the ABO. This suggests that the ABO V-ATPase is homologous to other known V-ATPase species. Finally, identification of V-ATPase was established phar- macologically using the specific V-ATPase inhibitor bafilo- mycin A, (Bowman et 1988). ATPase assays were conducted on the 1000 X g supernatant of ABO homoge- nates obtained from 15-20 animals. Three separate pooled ABO homogenates were produced and assayed for ATPase activity. Each 1000 X g supernatant provided sufficient protein to assay a minimum of three control and three inhibited trials. About 10% of the phosphate liberated in the control samples was seen to be derived from V-ATPase. V-ATPase activity ranged from to pmol Pi/(jug protein • min), with an average of ~67 pmolAjiig protein • min). The disparity in the ATPase activities among the three ABO homogenates can be attributed primarily to variation in the amount of V-ATPase actually present in each homog- enate (total protein was determined, so inclusion of excess foot tissue, for example, would lower the V-ATPase con- tent) and to variation in the amount of Pi released through dissociation of endogenous ATP or other
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
