PHD2基因在鲸类低氧信号通路中的功能

为研究鲸类低氧适应的分子机制,文章克隆了不同低氧耐受能力的3个鲸类物种,抹香鲸(Physeter macrocephalus)、白鲸(Delphinapterus leucas)和长江江豚(Neophocaena phocaenoids asiaeorientalis)的脯氨酸羟化酶2(PHD2)。通过对其序列进行分析,发现3个物种PHD2的氨基酸序列非常保守。通过对这3个物种的PHD2的功能进行探究发现:3个物种的PHD2在常氧情况下均可以降解3个物种的HIF-α(包括HIF-1α和HIF-2α)蛋白,而在低氧(O₂浓度小于2%)情况下,PHD2则无法明显降解HIF-α蛋白。在常氧下,鲸类的PHD2降解HIF-α是依赖于识别鲸类的HIF-1α上LTLLAP和LEMLAP,HIF-2α的LAQLAP和LETLAP氨基酸片段,推测PHD2是通过对HIF-α序列中的脯氨酸位点进行羟基化修饰后,被VHL-E3泛素连接酶复合体所识别,发生泛素化降解。而在低氧条件下,PHD2的活性受到抑制HIF-α不能被VHL-E3泛素连接酶复合体识别,发生降解。研究对3种不同低氧耐受能力...     

Ascorbate system in plant development

By using lycorine, a specific inhibitor of ascorbate biosynthesis, it was possible to demonstrate that plant cells consume a high quantity of ascorbate (AA). Thein vivo metabolic reactions utilizing ascorbate are the elimination of H₂O₂ by ascorbate peroxidase and the hydroxylation of proline residues present in the polypeptide chains by means of peptidyl-proline hydroxylase.Ascorbate acts in the cell metabolism as an electron donor, and consequently ascorbate free radical (AFR) is continuously produced. AFR can be reconverted to AA by means of AFR reductase or can undergo spontaneous disproportion, thus generating dehydroascorbic acid (DHA).During cell division and cell expansion ascorbate consumption is more or less the same; however, the AA/DHA ratio is 6–10 during cell division and 1–3 during cell expansion. This ratio depends essentially on the different AFR reductase activity in these cells. In meristematic cells AFR reductase is very high, and consequently a large amount of AFR is reduced to AA and a small amount of AFR undergoes disproportionation; in expanding cells the AFR reductase activity is lower, and therefore AFR is massively disproportionated, thus generating a large quantity of DHA. Since the transition from cell division to cell expansion is marked by a large drop of AFR reductase activity in the ER, it is suggested here that AFR formed in this compartment may be involved in the enlargement of the ER membranes and provacuole acidification.DHA is a toxic compound for the cell metabolism and as such the cell has various strategies to counteract its effects: (i) meristematic cells, having an elevated AFR reductase, prevent large DHA production, limiting the quantity of AFR undergoing disproportionation. (ii) Expanding cells, which contain a lower AFR reductase, are, however, provided with a developed vacuolar system and segregate the toxic DHA in the vacuole. (iii) Chloroplast strategy against DHA toxicity is efficient DHA reduction to AA using GSH as electron donor. This strategy is usually poorly utilized by the surrounding cytoplasm.DHA reduction does play an important role at one point in the life of the plant, that is, during the early stage of seed germination. The dry seed does not store ascorbate, but contains DHA, and several DHA-reducing proteins are detectable. In this condition, DHA reduction is necessary to form a limited AA pool in the seed for the metabolic requirements of the beginning of germination. After 30–40h ascorbateex novo synthesis starts, DHA reduction declines until a single isoform remains, as is typical in the roots, stem, and leaves of seedlings. Finally, DHA recycling also appears to be important under adverse environmental conditions and ascorbate deficiency.