Participation of Na,K-ATPase in FGF-2 secretion: Rescue of ouabain-inhibitable FGF-2 secretion by ouabain-resistant Na,K-ATPase α subunits

J. P. Dahl, A. Binda, Victor Canfield, Robert Levenson

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

Abstract

We have examined the relationship between Na,K-ATPase and FGF-2 secretion in transfected primate cells. FGF-2 lacks a classic hydrophobic export signal, and the mechanisms mediating its secretion are unknown. To monitor secretion, a FLAG epitope tag was inserted into the carboxyl terminus of the 18 kDa form of human FGF-2, and the construct was transfected into either human HEK 293 or monkey CV-1 cells. Exported FGF-2 was detected in the culture medium using the FLAG-specific monoclonal antibody M2. FGF-2 secretion from HEK 293 or CV-1 cells was linear over time and sensitive to inhibition by the cardiac glycoside ouabain, a specific inhibitor of the Na,K-ATPase. In contrast, the secretion of FGF-8 (an FGF family member that contains a hydrophobic secretory signal) was not inhibited by treatment of HEK 293 or CV-1 cells with ouabain. FGF-2 secretion was also assayed in CV-1 cells expressing the naturally ouabain-resistant rodent Na,K-ATPase α1 subunit. In cells expressing the rodent α1 subunit, FGF-2 secretion was unaffected by high levels of ouabain, indicating that the rodent α1 subunit was capable of rescuing ouabain-inhibitable FGF-2 export. Expression of ouabain-resistant mutants of the rodent α2 and α3 subunits, or the naturally ouabain-resistant rodent α4 subunit, also supported FGF-2 secretion in ouabain-treated cells. Taken together, our studies are consistent with the idea that the Na,K-ATPase plays a prominent role in regulating FGF-2 secretion, although none of the α subunit isoforms exhibited specificity with regard to FGF-2 export.

Original languageEnglish (US)
Pages (from-to)14877-14883
Number of pages7
JournalBiochemistry
Volume39
Issue number48
DOIs
StatePublished - Dec 5 2000

All Science Journal Classification (ASJC) codes

  • Biochemistry

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