Inhibitory effects of purified antibody against α-1 repeat(117-137)on Na+-Ca2+exchange and L-type Ca2+currents in rat cardiomyocytes

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FENG Qi-Long(Department of Physiology,Shanxi Medical University,Taiyuan,030001,China;Key Laboratory of Cellular Physiology,Shanxi Province and Ministry of Education,Taiyuan 030001,China)
WU Dong-Mei(Department of Pharmacology,Shanxi Medical University,Taiyuan,030001,China;Key Laboratory of Cellular Physiology,Shanxi Province and Ministry of Education,Taiyuan 030001,China)
CUI Xiang-Li(Department of Physiology,Shanxi Medical University,Taiyuan,030001,China;Key Laboratory of Cellular Physiology,Shanxi Province and Ministry of Education,Taiyuan 030001,China)
ZHAO Hua-Chen(Beijing Cord Blood Bank,Beijing 100176,China)
LIN Yuan-Yuan(Department of Physiology,Shanxi Medical University,Taiyuan,030001,China)
ZHAO Lu-Ying(Department of Physiology,Shanxi Medical University,Taiyuan,030001,China)
WU Bo-Wei(Department of Physiology,Shanxi Medical University,Taiyuan,030001,China;Key Laboratory of Cellular Physiology,Shanxi Province and Ministry of Education,Taiyuan 030001,China)
Journal Title:
Volume 62, Issue 05, 2010
Key Word:
sodium-calcium exchanger;patch-clamp technique;cardiac myocytes;antibody

Abstract: Considering that α-1 repeat region may be involved in the ion binding and translocation of Na+-Ca2+exchanger(NCX),it is possible that the antibodies against NCX a-1 repeat may have a crucial action on NCX activity.The aim of the present study is to investigate the effect of antibody against α-1 repeat(117-137),designated as α-1(117-137),on NCX activity.The antibody against the synthesized α-1(117-137)was prepared and affinity-purified.Whole-cell patch clamp technique was used to study the change of Na+-Ca2+exchange current(INa/Ca)in adult rat cardiomyocytes.To evaluate the functional specificity of this antibody,its effects on L-type Ca2+current(ICa,L),voltage-gated Na+current(INa)and delayed rectifier K+current(IK)were also observed.The amino acid sequences ofα-1(117-137)in NCX and residues 1 076-1 096 within L-type Ca2+channel were compared using EMBOSS Pairwise Alignment Algorithms.The results showed that outward and inward INa/Ca were decreased by the antibody against α-1(117-137)dose-dependently in the concentration range from 10 to 160 nmol/L,with IC50 values of 18.9 nmol/L and 22.4 nmol/L,respectively.Meanwhile,the antibody also decreased a concentration-dependent manner with IC50 of 22.7 nmol/L.No obvious effects of the antibody on INa and IK were observed.Moreover,comparison of the amino acid sequences showed there was 23.8% sequence similarity between NCXα-1(117-137)and residues 1 076-1 096 within L-type Ca2+channel.These results suggest that antibody against α-1(117-137)is a blocking antibody to NCX and can also decrease ICa,L in a concentration-dependent manner,while it does not have obvious effects on INa and IK.

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