Functional interdependence and colocalization of endothelial nitric oxide synthase and heat shock protein 90 in cerebral arteries

Vini G. Khurana, Kristian Feterik, Margaret J. Springett, Daihiko Eguchi, Vijay Shah, Zvonimir S Katusic

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Heat shock protein 90 (HSP90), an essential component of several signal transduction systems, participates in the activation of endothelial nitric oxide synthase (eNOS) in cells. The objective of the current study was to determine if HSP90 and eNOS were functionally interdependent and colocalized in the cerebral circulation. The authors used isometric force recording, cyclic 3'5'-guanosine monophosphate (cGMP) radioimmunoassay (RIA), and immunogold electron microscopy (EM) to study canine basilar artery. They found that geldanamycin (0.1 to 10 μg/mL), a selective HSP90 inhibitor, caused concentration-dependent contractions in arterial rings (n = 6 dogs). Contractions to geldanamycin were unaffected by a cyclooxygenase inhibitor, indomethacin (10 μmol/L; P < 0.05, n = 6). Functional evidence for interaction between HSP90 and nitric oxide (NO)-mediated signaling included observations that the contractile effect of geldanamycin was the following: (1) endothelium-dependent, (2) abolished by N(g)-nitro-L-arginine methylester (L-NAME; 0.3 mmol/L), and (3) nonadditive with the contractile effect of this NOS inhibitor (P <0.01, n = 6 for each). Furthermore, RIA showed significant reduction in cGMP levels in arteries treated with geldanamycin (3 μg/mL; P < 0.02, n = 8), whereas immunogold EM demonstrated areas of colocalization of HSP90 and eNOS selectively in the cytoplasm of endothelial cells. The current findings suggest that in cerebral arteries, endothelial HSP90 plays an important role in modulation of basal NO-mediated signaling. This interaction may be particularly important in stress-induced up-regulation of HSP90 with subsequent alteration of vasomotor function.

Original languageEnglish (US)
Pages (from-to)1563-1570
Number of pages8
JournalJournal of Cerebral Blood Flow and Metabolism
Volume20
Issue number11
StatePublished - 2000

Fingerprint

HSP90 Heat-Shock Proteins
Cerebral Arteries
Nitric Oxide Synthase Type III
Radioimmunoassay
Cerebrovascular Circulation
Electron Microscopy
Nitric Oxide
Basilar Artery
Cyclooxygenase Inhibitors
NG-Nitroarginine Methyl Ester
Indomethacin
Endothelium
Arginine
Canidae
Signal Transduction
Cytoplasm
Up-Regulation
Endothelial Cells
Arteries
Dogs

Keywords

  • Endothelium
  • Heat shock protein
  • HSP90
  • Nitric oxide synthase

ASJC Scopus subject areas

  • Endocrinology
  • Neuroscience(all)
  • Endocrinology, Diabetes and Metabolism

Cite this

Functional interdependence and colocalization of endothelial nitric oxide synthase and heat shock protein 90 in cerebral arteries. / Khurana, Vini G.; Feterik, Kristian; Springett, Margaret J.; Eguchi, Daihiko; Shah, Vijay; Katusic, Zvonimir S.

In: Journal of Cerebral Blood Flow and Metabolism, Vol. 20, No. 11, 2000, p. 1563-1570.

Research output: Contribution to journalArticle

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AU - Shah, Vijay

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