Hazelton G A, Lang C A
Biochem J. 1983 Feb 15;210(2):289-95. doi: 10.1042/bj2100289.
Our previous findings [Hazelton & Lang (1978) Fed. Proc. Fed. Am. Soc. Exp. Biol. 37(6), 2378 (abstr.)] demonstrated a senescence-specific decrease in glutathione (GSH) concentration in the yellow-fever mosquito Aedes aegypti (Louisville)]. As a possible mechanism for this change, GSH biosynthesis was investigated in adult mosquitoes of different ages through the life-span. Biosynthesis was measured as the incorporation rate of [14C]glycine or [14C]cystine into glutathione. Essential information to validate the procedure was also obtained on the precursor-amino-acid pool sizes and kinetic parameters such as lag-time and time course of incorporation. Also, synthesis de novo rather than exchange was verified using buthionine sulphoximine, a specific inhibitor of GSH biosynthesis. The synthetic rates with either amino acid precursor varied throughout the adult life-span, but the patterns for both precursors were essentially identical. Biosynthesis was high in the newly emerged adult and decreased 62-70% (P less than 0.005) to a plateau during maturity. From the mature value there was a decrease of 36-41% (P less than 0.005) to a new plateau during senescence. Glutathione biosynthesis and concentration were correlated throughout maturity and senescence (r = 0.982) and thus biosynthesis was proportional to glutathione content. On this basis we concluded that impaired biosynthesis is the major and perhaps sole mechanism for the aging decrease in glutathione content.
我们之前的研究结果[黑泽尔顿和朗(1978年),《联邦程序汇编》,美国实验生物学联合会会刊37(6),2378(摘要)]表明,黄热病蚊子埃及伊蚊(路易斯维尔种群)体内的谷胱甘肽(GSH)浓度会出现与衰老相关的特异性下降。作为这种变化的一种可能机制,我们对不同年龄的成年蚊子在其整个生命周期内的GSH生物合成进行了研究。生物合成通过[14C]甘氨酸或[14C]胱氨酸掺入谷胱甘肽的速率来衡量。我们还获得了验证该程序所需的重要信息,包括前体氨基酸库大小以及诸如滞后时间和掺入时间进程等动力学参数。此外,使用谷胱甘肽生物合成的特异性抑制剂丁硫氨酸亚砜胺验证了从头合成而非交换过程。两种氨基酸前体的合成速率在成年蚊子的整个生命周期中都有所变化,但两种前体的模式基本相同。新羽化的成虫中生物合成水平较高,在成熟阶段下降了62 - 70%(P < 0.005),并达到一个稳定水平。从成熟阶段的水平开始,在衰老阶段又下降了36 - 41%(P < 0.005),达到一个新的稳定水平。在整个成熟和衰老阶段,谷胱甘肽生物合成与浓度呈正相关(r = 0.982),因此生物合成与谷胱甘肽含量成正比。基于此,我们得出结论,生物合成受损是谷胱甘肽含量随衰老而下降的主要且可能是唯一的机制。