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Inhibition of Na-K-Cl cotransport in Ehrlich ascites cells by antiserum against purified proteins of the cotransporter

Journal Article · · Proceedings of the National Academy of Sciences of the United States of America; (United States)
 [1]; ;  [2]
  1. Syracuse Univ., NY (USA)
  2. Univ. of Copenhagen (Denmark)

Two proteins were purified earlier from solubilized membranes of Ehrlich ascites cells by using a bumetanide-Sepharose affinity column. These proteins were proposed to be constituents of the Na-K-Cl cotransporter. However, the specificity of binding of bumetanide to the cotransporter was insufficient evidence for this proposal. The authors now have direct evidence that the purified protein contains components of the cotransporter. Antiserum raised against the bumetanide-binding proteins strongly inhibits Na-K-Cl cotransport measured by two independent methods. Cotransport was induced by hypertonic challenge and was measured as the bumetanide-sensitive portion of unidirectional Cl influx and as regulatory cell volume increase. In both assays, contransport was strongly inhibited by antiserum. Fab fragments of the antibodies inhibited cotransport to a similar extent.

OSTI ID:
5452080
Journal Information:
Proceedings of the National Academy of Sciences of the United States of America; (United States), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (United States) Vol. 87:17; ISSN PNASA; ISSN 0027-8424
Country of Publication:
United States
Language:
English