Brahm J
J Gen Physiol. 1977 Sep;70(3):283-306. doi: 10.1085/jgp.70.3.283.
Chloride self-exchange in human red cells was studied between 0 degrees C and 38 degrees C. At higher temperatures the flow-tube method was used. Although the general features of chloride transport at 0 degrees C and 38 degrees C are similar, the following differences were found: (a) the maximum pH of chloride self-exchange flux was lowered 0.6 pH unit from 7.8 to 7.2 when temperature was increased from 0 degrees C to 38 degrees C; (b)the apparent half-saturation constant increased from 28 mM at 0 degrees C to 65 mM at 38 degrees C; (c) chloride transport at body temperature is slower than predicted by other investigators by extrapolation from low-temperature results. Chloride transport increased only 200 times when temperature was raised from 0 degrees C to 38 degrees C, because the apparent activation energy decreased from 30 kcal mol(-1) to 20 kcal mol(-1) above a temperature of 15 degrees C; (d) a study of temperature dependence of the slower bromide self-exchange showed that a similar change of activation energy occurred around 25 degrees C. Both in the case of Cl(-) (15 degrees C) and in the case of Br(-) (25 degrees C), critical temperature was reached when the anion self-exchange had a turnover number of about 4x10(9) ions cell (-1)s(-1); (e) inhibition of chloride transport by DIDS (4,4'- diisothiocyano-stilbene-2,2'-disulfonate)revealed that the deflection persisted at 15 degrees C at partial inhibition (66 percent) presumably because DIDS inactivated 66 percent of the transport sites. It is suggested that a less temperature- dependent step of anion exchange becomes rate limiting at the temperature where a critical turnover number is reached.
在0℃至38℃之间研究了人体红细胞中的氯离子自我交换。在较高温度下使用流动管法。尽管0℃和38℃时氯离子转运的总体特征相似,但发现了以下差异:(a)当温度从0℃升至38℃时,氯离子自我交换通量的最大pH值从7.8降低了0.6个pH单位至7.2;(b)表观半饱和常数从0℃时的28 mM增加到38℃时的65 mM;(c)体温下的氯离子转运比其他研究者根据低温结果外推预测的要慢。当温度从0℃升至38℃时,氯离子转运仅增加了200倍,因为在15℃以上表观活化能从30 kcal mol⁻¹降至20 kcal mol⁻¹;(d)对较慢的溴离子自我交换的温度依赖性研究表明,在25℃左右发生了类似的活化能变化。无论是Cl⁻(15℃)还是Br⁻(25℃)的情况,当阴离子自我交换的周转数约为4×10⁹离子·细胞⁻¹·秒⁻¹时达到临界温度;(e)DIDS(4,4'-二异硫氰基芪-2,2'-二磺酸盐)对氯离子转运的抑制作用表明,在15℃下部分抑制(66%)时偏差仍然存在,大概是因为DIDS使66%的转运位点失活。有人提出,在达到临界周转数的温度下,阴离子交换中一个对温度依赖性较小的步骤成为限速步骤。