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Youngsuk Lee,Nari Kim,Hyunju Kim,Hyun Joo,Youngnam Kim,Daehoon Jeong,Dang Van Cuong,Euiyong Kim,Dae Young Hur,Young Shik Park,Yong Geun Hong,Sangkyung Lee,Joonyong Chung,Daehyun Seog,Jin Han 대한생리학회-대한약리학회 2004 The Korean Journal of Physiology & Pharmacology Vol.8 No.4
The purpose of the present study was to evaluate the expression of cardiac marker protein in rabbit cardiac tissue that was exposed to ischemic preconditioning (IPC), or ischemiareperfusion injury (IR) using two-dimensional gel electrophoresis (2DE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). We compared 2DE gels of control (uninjured) cardiac tissue with those of IPC and IR cardiac tissue. Expression of one protein was detected in IR heart tissue, however the protein was not detected in the samples of control and IPC tissue. To further characterize the detected protein molecule, the protein in the 2D gel was isolated and subjected to trypsin digestion, followed by MALDI-MS. The protein was identified as myoglobin, which was confirmed also by Western blot analysis. These results are consistent with previous studies of cardiac markers in ischemic hearts, indicating myoglobin as a suitable marker of myocardial injury. In addition, the present use of multiple techniques indicates that proteomic analysis is an appropriate means to identify cardiac markers in studies of IPC and IR.
Detection of Mitochondrial ATP-Sensitive Potassium Channels in Rat Cardiomyocytes
Dang Van Cuong,Nari Kim,Euiyong Kim,Youngsuk Lee,Hyunju Kim,Sunghyun Kang,Dae Young Hur,Hyun Joo,Young Shik Park,Yong Geun Hong,Sangkyung Lee,Joonyong Chung,Daehyun Seog,Jin Han 대한생리학회-대한약리학회 2004 The Korean Journal of Physiology & Pharmacology Vol.8 No.4
Mitochondrial ATP-sensitive potassium (mitoK<SUB>ATP</SUB>) channels play a role in early and late ischemic preconditioning. Nevertheless, the subunit composition of mitoK<SUB>ATP</SUB> channels remains unclear. In this study, we investigated the subunit composition of mitoK<SUB>ATP</SUB> channels in mitochondria isolated from rat cardiac<FONT COLOR=RED><SPAN STYLE= font-size:11pt; > </SPAN>myocytes. Mitochondria were visualized using the red fluorescence probe, Mitrotracker Red, while mitoK<SUB>ATP</SUB> channels were visualized using the green fluorescence probe, glibenclamide-BODIPY. The immunofluorescence confocal microscopy revealed the presence of Kir6.1, Kir6.2 and SUR2 present in the cardiac mitochondria. Western blot analysis was carried to further investigate the nature of mitoK<SUB>ATP</SUB> channels. For SUR proteins, a 140-kDa immunoreactive band that corresponded to SUR2, but no SUR1 was detected. For Kir6.2, three bands (∼44, ∼46, and ∼30 kDa) were detected, and a specific ∼46-kDa immunoreactive band corresponding to Kir6.1 was also observed. These observations suggest that the subunits of mitoK<SUB>ATP</SUB> channels in rat myocytes include Kir6.1, Kir6.2, and a<FONT COLOR=RED><SPAN STYLE= font-size:11pt; > </SPAN>SUR2-related sulfonylurea-binding protein.