Korean J Physiol Pharmacol.  2004 Aug;8(4):207-211.

Identification of a Marker Protein for Cardiac Ischemia and Reperfusion Injury by Two-Dimensional Gel Electrophoresis and Matrix-Assisted Laser Desorption Ionization Mass Spectrometry

• Lee Y
• Kim N
• Kim H
• Joo H
• Kim Y
• Jeong D
• Cuong DV
• Kim E
• Hur DY
• Park YS
• Hong YG
• Lee S
• Chung J
• Seog D
• Han J

Abstract

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.

Keyword

Myoglobin; Ischemiareperfusion injury; Cardiac injury marker; Two-dimensional gel electrophoresis; Matrix-assisted laser desorption ionization mass spectrometry

MeSH Terms

Blotting, Western
Digestion
Electrophoresis, Gel, Two-Dimensional*
Gels
Heart
Ischemia*
Ischemic Preconditioning
Mass Spectrometry*
Myoglobin
Reperfusion Injury*
Reperfusion*
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Trypsin
Gels
Myoglobin
Trypsin