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Characterization of the placental brush border membrane Na+/H+ exchanger: identification of thiol-dependent transitions in apparent molecular size.

ABSTRACT: We examined the protein and mRNA encoding the amiloride-sensitive Na+/H+ exchanger from human placenta. Reverse transcriptase PCR of human placental RNA and a human choriocarcinoma cell line showed that the message for the amiloride-sensitive Na+/H+ exchanger from human placenta. Reverse transcriptase PCR of human placental RNA and a human choriocarcinoma cell line showed that the message for the amiloride-sensitive Na+/H+ exchanger is present in the placenta and its derived cell line. Northern blot analysis showed only one species of Na+/H+ exchanger mRNA, of about 5 kb in size. To examine the Na+/H+ exchanger protein two different affinity-purified antibodies were produced against the C-terminal cytoplasmic region of the Na+/H+ exchanger. The antibodies both identified a 105 kDa protein in human placental brush border membrane vesicles. Under non-reducing conditions the amount of 105 kDa protein was greatly decreased, while a 205 kDa protein became apparent. This is probably a dimer of the 105 kDa protein. The monomer-to-dimer transition was dependent on the concentration of beta-mercaptoethanol. The results show that the amiloride-sensitive Na+/H+ exchanger is relatively abundant in human placenta and that it can exist as a larger 205 kDa protein linked by disulphide bonds.

SUBMITTER: Fliegel L

PROVIDER: S-EPMC1132136 | biostudies-other | 1993 Jan

REPOSITORIES: biostudies-other

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Sodium-gradient-driven, high-affinity, uphill transport of succinate in human placental brush-border membrane vesicles.

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