The GSTM2 C-Terminal Domain Depresses Contractility and Ca2+ Transients in Neonatal Rat Ventricular Cardiomyocytes.
The cardiac ryanodine receptor (RyR2) is an intracellular ion channel that regulates Ca2+ release from the sarcoplasmic reticulum (SR) during excitation-contraction coupling in the heart. The glutathione transferases (GSTs) are a family of phase II detoxification enzymes with additional functions including the selective inhibition of RyR2, with therapeutic implications. The C-terminal half of GSTM2 (GSTM2C) is essential for RyR2 inhibition, and mutations F157A and Y160A within GSTM2C prevent the inhibitory action. Our objective in this investigation was to determine whether GSTM2C can enter cultured rat neonatal ventricular cardiomyocytes and influence contractility. We show that oregon green-tagged GSTM2C (at 1 μM) is internalized into the myocytes and it reduces spontaneous contraction frequency and myocyte shortening. Field stimulation of myocytes evoked contraction in the same percentage of myocytes treated either with media alone or media plus 15 μM GSTM2C. Myocyte shortening during contraction was significantly reduced by exposure to 15 μM GSTM2C, but not 5 and 10 μM GSTM2C and was unaffected by exposure to 15 μM of the mutants Y160A or F157A. The amplitude of the Ca2+ transient in the 15 μM GSTM2C - treated myocytes was significantly decreased, the rise time was significantly longer and the decay time was significantly shorter than in control myocytes. The Ca2+ transient was not altered by exposure to Y160A or F157A. The results are consistent with GSTM2C entering the myocytes and inhibiting RyR2, in a manner that indicates a possible therapeutic potential for treatment of arrhythmia in the neonatal heart.
John Curtin School of Medical Research, The Australian National University, GPO Box 334, Canberra City, ACT 2600, Australia.
Ruwani P
RP
Hewawasam
John Curtin School of Medical Research, The Australian National University, GPO Box 334, Canberra City, ACT 2600, Australia.
Dan
D
Liu
John Curtin School of Medical Research, The Australian National University, GPO Box 334, Canberra City, ACT 2600, Australia.
Marco G
MG
Casarotto
John Curtin School of Medical Research, The Australian National University, GPO Box 334, Canberra City, ACT 2600, Australia.
Philip G
PG
Board
John Curtin School of Medical Research, The Australian National University, GPO Box 334, Canberra City, ACT 2600, Australia.
Angela F
AF
Dulhunty
D000185:Actinin; D000818:Animals; D000831:Animals, Newborn; D020013:Calcium Signaling; D004558:Electric Stimulation; D005982:Glutathione Transferase / Q000737:chemistry / Q000378:metabolism*; D006352:Heart Ventricles / Q000378:metabolism*; D008958:Models, Molecular; D009200:Myocardial Contraction; D032383:Myocytes, Cardiac / Q000378:metabolism*; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D051381:Rats
D000185:Actinin; D005982:Glutathione Transferase; C412602:glutathione S-transferase Mu 2