Direct interaction of the cell division cycle 37 homolog inhibits endothelial nitric oxide synthase activity

M. Brennan Harris, Manuela Bartoli, Sarika G. Sood, Robert L. Matts, Richard C Venema

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Endothelial NO synthase (eNOS) via the production of NO in the endothelium plays a key role in cardiovascular biology and is tightly regulated by co- and posttranslational mechanisms, phosphorylation, and protein-protein interactions. The cell division cycle 37 homolog (Cdc37) is a key heat shock protein 90 (Hsp90) cochaperone for protein kinase clients, and Akt/Hsp90 interaction is dependent on Cdc37. Because both Hsp90 and Akt are key eNOS regulatory proteins, we hypothesized that Cdc37 interacts with eNOS as part of the regulatory complex. In the present study, we demonstrate by coimmunoprecipitation and affinity purification in bovine aortic endothelial cells (BAECs) that Cdc37 is complexed with eNOS, Hsp90, and Akt. In addition, cell fractionation data indicate that Cdc37 is found in caveolae with eNOS. Further analysis by in vitro binding assays reveals a direct interaction between purified Cdc37 and eNOS. Incubation of purified Cdc37 with purified wild-type eNOS decreases eNOS activity in vitro. Overexpression of wild-type Cdc37 in BAECs inhibits eNOS activity and NO release, whereas overexpression of S13A-Cdc37 mutant in BAECs increases eNOS activity and NO release. Taken together, these data suggest that Cdc37 has a direct regulatory interaction with eNOS and may play an important role in mediating the eNOS protein complex formation as well as subsequent eNOS phosphorylation and activation.

Original languageEnglish (US)
Pages (from-to)335-341
Number of pages7
JournalCirculation research
Volume98
Issue number3
DOIs
StatePublished - Feb 1 2006

Fingerprint

Nitric Oxide Synthase Type III
Nitric Oxide Synthase
Cell Cycle
HSP90 Heat-Shock Proteins
Endothelial Cells
Phosphorylation
Cell Fractionation
Caveolae
Cell Cycle Proteins
Proteins
Protein Kinases
Endothelium

Keywords

  • Cdc37
  • Endothelium
  • Heat shock protein 90
  • Nitric oxide synthase

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Direct interaction of the cell division cycle 37 homolog inhibits endothelial nitric oxide synthase activity. / Harris, M. Brennan; Bartoli, Manuela; Sood, Sarika G.; Matts, Robert L.; Venema, Richard C.

In: Circulation research, Vol. 98, No. 3, 01.02.2006, p. 335-341.

Research output: Contribution to journalArticle

Harris, M. Brennan ; Bartoli, Manuela ; Sood, Sarika G. ; Matts, Robert L. ; Venema, Richard C. / Direct interaction of the cell division cycle 37 homolog inhibits endothelial nitric oxide synthase activity. In: Circulation research. 2006 ; Vol. 98, No. 3. pp. 335-341.
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