Hainsworth A H, Levis R A, Eisenberg R S
Department of Molecular Biophysics and Physiology, Rush Medical Center, Chicago, Illinois 60612.
J Gen Physiol. 1994 Nov;104(5):857-83. doi: 10.1085/jgp.104.5.857.
Open-channel noise was studied in the large potassium channel of the sarcoplasmic reticulum (SR). Inside-out patches were excised directly from the SR of split skeletal muscle fibers of lobster, with lobster relaxing ringer (LRR) in bath and pipette. The power spectrum of open-channel noise is very low and approximately flat in the 100 Hz-10 kHz frequency range. At 20 degrees C, with an applied voltage of 50 mV, the mean single-channel current (i) is 9 pA (mean single-channel conductance = 180 pS) and the mean power spectral density 1.1 x 10(-29) A2/Hz. The latter increases nonlinearly with (i), showing a progressively steeper dependence as (i) increases. At 20 mV, the mean power spectral density is almost independent of (i) and approximately 1.4 times that of the Johnson noise calculated for the equivalent ideal resistor with zero net current; at 70 mV it increases approximately in proportion to (i)2. The mean power spectral density has a weak temperature dependence, very similar to that of (i), and both are well described by a Q10 of 1.3 throughout the range 3-40 degrees C. Discrete ion transport events are thought to account for a significant fraction of the measured open-channel noise, probably approximately 30-50% at 50 mV. Brief interruptions of the single-channel current, due either to blockage of the open channel by an extrinsic aqueous species, or to intrinsic conformational changes in the channel molecule itself, were a possible additional source of open-channel noise. Experiments in modified bathing solutions indicate, however, that open-channel noise is not affected by any of the identified aqueous species present in LRR. In particular, magnesium ions, the species thought most likely to cause brief blockages, and calcium and hydrogen ions, have no detectable effect. This channel's openings exhibit many brief closings and substrates, due to intrinsic gating of the channel. Unresolved brief full closings are calculated to make a negligible contribution (< 1%) to the measured power spectral density. The only significant source of noise due to band width-limited missed events is brief, frequent 80% substrates (mean duration 20 microseconds, mean frequency 1,000 s-1) which account for a small part of the measured power spectral density (approximately 14%, at 50 mV, 20 degrees C). We conclude that a large fraction of the measured open-channel noise results from intrinsic conductance fluctuations, with a corner frequency higher than the resolution of our recordings, in the range 10(4)-10(7) Hz.(ABSTRACT TRUNCATED AT 400 WORDS)
对肌浆网(SR)大钾通道的明渠噪声进行了研究。内面向外的膜片直接从龙虾分裂骨骼肌纤维的SR中切除,浴槽和移液管中使用龙虾松弛林格液(LRR)。在100 Hz - 10 kHz频率范围内,明渠噪声的功率谱非常低且近似平坦。在20摄氏度、施加电压为50 mV时,平均单通道电流(i)为9 pA(平均单通道电导 = 180 pS),平均功率谱密度为1.1×10⁻²⁹ A²/Hz。后者随(i)非线性增加,随着(i)增大依赖性逐渐变陡。在20 mV时,平均功率谱密度几乎与(i)无关,约为等效零净电流理想电阻计算出的约翰逊噪声的1.4倍;在70 mV时,它大致与(i)²成比例增加。平均功率谱密度对温度的依赖性较弱,与(i)的依赖性非常相似,在3 - 40摄氏度范围内二者均可用1.3的Q₁₀很好地描述。离散离子转运事件被认为占测量到的明渠噪声的很大一部分,在50 mV时可能约为30 - 50%。单通道电流的短暂中断,要么是由于外部水性物质堵塞开放通道,要么是由于通道分子本身的内在构象变化,可能是明渠噪声的另一个来源。然而,在改良浴液中的实验表明,明渠噪声不受LRR中存在的任何已确定水性物质的影响。特别是,最有可能导致短暂堵塞的镁离子以及钙和氢离子均无可检测到的影响。由于通道的内在门控作用,该通道的开放表现出许多短暂关闭和底物。计算得出未解决的短暂完全关闭对测量到的功率谱密度的贡献可忽略不计(<1%)。由于带宽限制的漏检事件导致的唯一显著噪声源是短暂、频繁的80%底物(平均持续时间20微秒,平均频率1000 s⁻¹),它们占测量到的功率谱密度的一小部分(在50 mV、20摄氏度时约为14%)。我们得出结论,测量到的明渠噪声的很大一部分源于内在电导波动,其转折频率高于我们记录的分辨率,在10⁴ - 10⁷ Hz范围内。(摘要截于400字)