Phosphorylation of the heat shock-related protein, HSP20, mediates cyclic nucleotide-dependent relaxation

David A Woodrum, Walter Pipkin, Deron Tessier, Padmini Komalavilas, Colleen M. Brophy

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Cyclic nucleotide-dependent relaxation of vascular smooth muscle is associated with increases in the phosphorylation of the small heat shock-related protein, HSP20. To determine whether phosphorylated HSP20 directly mediates relaxation, we used gene transfection and protein transduction of HSP20 analogues. Rat mesangial cells were transfected with constructs containing wild-type HSP20-enhanced green fluorescent protein (EGFP), phosphorylation site mutated HSP20 (S16A-HSP20-EGFP), or EGFP alone. Contractile properties were determined on a silicone polymer substrata. In the presence of serum, EGFP-vector transfected control cells and S16A-HSP20 transfected cells formed wrinkles on the polymer (contracted). Activation of cyclic nucleotide signaling pathways in the EGFP-vector transfected control cells led to a time-dependent decrease in the wrinkles (relaxation). The S16A-HSP20 transfected cells were refractory to cyclic nucleotide-dependent relaxation. Cells overexpressing the wild-type HSP20 did not form wrinkles on the polymer in response to serum (refractory to contraction). Treatment of precontracted strips of intact bovine carotid artery smooth muscle with synthetic peptides containing HIV-trans-activating transcriptional activator and a phosphopeptide motif of HSP20 led to dose-dependent relaxation. These data provide evidence that phosphorylated HSP20 has a direct role in smooth muscle relaxation and that small phosphopeptide motifs of HSP20 can mimic the effects of the entire molecule.

Original languageEnglish (US)
Pages (from-to)874-881
Number of pages8
JournalJournal of Vascular Surgery
Volume37
Issue number4
DOIs
StatePublished - Apr 1 2003

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HSP20 Heat-Shock Proteins
Cyclic Nucleotides
Phosphorylation
Phosphopeptides
Polymers
Smooth Muscle
Small Heat-Shock Proteins
Muscle Relaxation
Mesangial Cells
Silicones
Serum
Vascular Smooth Muscle
Carotid Arteries
Transfection
enhanced green fluorescent protein
HIV
Peptides

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Surgery

Cite this

Phosphorylation of the heat shock-related protein, HSP20, mediates cyclic nucleotide-dependent relaxation. / Woodrum, David A; Pipkin, Walter; Tessier, Deron; Komalavilas, Padmini; Brophy, Colleen M.

In: Journal of Vascular Surgery, Vol. 37, No. 4, 01.04.2003, p. 874-881.

Research output: Contribution to journalArticle

Woodrum, David A ; Pipkin, Walter ; Tessier, Deron ; Komalavilas, Padmini ; Brophy, Colleen M. / Phosphorylation of the heat shock-related protein, HSP20, mediates cyclic nucleotide-dependent relaxation. In: Journal of Vascular Surgery. 2003 ; Vol. 37, No. 4. pp. 874-881.
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