Heat shock augments myosin phosphatase target-subunit phosphorylation

Abstract

Our previous study demonstrated that heat shock augmented vascular contraction. In the present study, we hypothesized that heat shock augments myosin phosphatase target-subunit (MYPT1) phosphorylation resulting in augmented vascular contraction. Endothelium-denuded rat aortic rings were mounted in organ baths, exposed to heat shock (42 °C for 45 min), and subjected to contraction 4 h after the heat shock followed by Western blot analysis for MLC 20 (the 20 kDa light chains of myosin II) or MYPT1. The contractile responses in both control and heat shock-treated aorta were inhibited by Y27632, an inhibitor of Rho-kinase. The level of the MLC 20 and MYPT1 Thr855 phosphorylation in response to KCl was higher in heat shock-treated aorta than that in timed-control. The increased MYPT1 Thr855 phosphorylation was inhibited by Y27632 (1.0 μM) in parallel with inhibition of MLC 20 phosphorylation and vascular contraction. These results indicate that heat shock augments MYPT1 phosphorylation resulting in augmented vascular contraction.