Molecular nature of sulfhydryl modification by hydrogen peroxide on type 1 ryanodine receptor.
OBJECTIVE
To elucidate the molecular nature of sulfhydryl modification by hydrogen peroxide on type 1 ryanodine receptor (RyR1).


METHODS
Rabbit skeletal muscle sarcoplasmic reticulum was treated with hydrogen peroxide, then RyR1 complex was isolated. The proteins in the complex were analysed by electrophoresis, Western blot and electron microscopy.


RESULTS
(1) Hydrogen peroxide induces inter-subunit cross-linking within the tetrameric RyR1 molecule; (2) in parallel to inter-subunit cross-linking, the RyR1 molecule changes morphology; (3) the chemical and morphological changes are reversible: upon reduction by reducing agents, the RyR1 molecule regains its original state.


CONCLUSIONS
These findings suggest that the molecular mechanism of RyR1 channel activity in sarcoplasmic reticulum regulated by hydrogen peroxide is through inter-subunit cross-linking within the tetrameric RyR1 molecule, which in turn induces structural changes of RyR1.
Han
Hong-mei
HM
Department of Biophysics, Peking University Health Science Centre, Peking University, Beijing 100083, China.
Wei
Ri-sheng
RS
Lai
F Anthony
FA
Lai
Anthony F
AF
Yin
Chang-cheng
CC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D022162:Cytoplasmic Vesicles / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D009124:Muscle Proteins / Q000378:metabolism*; D016877:Oxidants / Q000494:pharmacology; D011817:Rabbits; D012433:Ryanodine / Q000378:metabolism; D019837:Ryanodine Receptor Calcium Release Channel / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D012519:Sarcoplasmic Reticulum / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D009124:Muscle Proteins; D016877:Oxidants; D019837:Ryanodine Receptor Calcium Release Channel; D013438:Sulfhydryl Compounds; D012433:Ryanodine; D006861:Hydrogen Peroxide; D002118:Calcium