Kameyama M, Kameyama A, Nakayama T, Kaibara M
National Institute for Physiological Sciences, Okazaki, Japan.
Pflugers Arch. 1988 Aug;412(3):328-30. doi: 10.1007/BF00582516.
Effects of cardiac-tissue extract on the activity of L-type Ca2+ channels were investigated in guinea-pig ventricular myocytes with the patch-clamp method. In most patches, Ca2+-channel current recorded with a pipette solution containing 50 mM Ba2+ and 3 microM Bay K 8644 ran down within 5 min after excision of the patches into a solution containing EGTA. This run-down of Ca2+ channels was prevented when patches were excised into a solution containing a supernatant fraction of homogenate of guinea-pig or bovine heart. Furthermore, this tissue extract was able to restore channel activity after run-down. This channel-activating effect of the extract was abolished by heat treatment or trypsin digestion. Fractionation of the extract by gel filtration suggested that the channel-activating factor(s) had an apparent molecular weight of 2-3 x 10(5). These results suggest that some cytoplasmic protein(s) maintains the activity of the cardiac L-type Ca2+ channel.
采用膜片钳技术,在豚鼠心室肌细胞中研究了心脏组织提取物对L型Ca2+通道活性的影响。在大多数膜片中,用含50 mM Ba2+和3 microM Bay K 8644的移液管溶液记录的Ca2+通道电流,在膜片切除到含EGTA的溶液中后5分钟内就衰减了。当膜片切除到含豚鼠或牛心脏匀浆上清液的溶液中时,Ca2+通道的这种衰减被阻止。此外,这种组织提取物能够在衰减后恢复通道活性。提取物的这种通道激活作用被热处理或胰蛋白酶消化所消除。通过凝胶过滤对提取物进行分级分离表明,通道激活因子的表观分子量为2 - 3×10(5)。这些结果表明,某些细胞质蛋白维持心脏L型Ca2+通道的活性。