Proteomic characterization of the human sperm nucleus
Proteomic characterization of the human sperm nucleus
复制标题
DOI：
10.1002/pmic.201000799
复制
发表时间：
2011-07-01
期刊：
影响因子：
3.4
通讯作者：
Oliva, Rafael
中科院分区：
文献类型：
--
作者：
de Mateo, Sara;Castillo, Judit;Oliva, Rafael
生成精子核蛋白目录是阐明受精时传递给卵母细胞的父本染色质功能的重要的第一步。为了实现这一目标，通过 CTAB 处理获得了精子核，并通过光学显微镜和透射电子显微镜评估，其纯度超过 99.9%，不含任何尾部碎片、顶体或线粒体。提取核蛋白并在 2-D 和 1-D 凝胶中分离，并切除 2-D 斑点和 1-D 带并进行分析，以通过 LC-MS/MS 鉴定蛋白质。通过这种方法，从分离的精子核中鉴定出了 403 种不同的蛋白质。已鉴定的最丰富的蛋白质家族是组蛋白，其一些新成员以前从未报道过存在于生精细胞系或人类成熟精子中。超过一半（52.6%）的蛋白质在之前的人类全精子细胞蛋白质组报告中未被检测到。相关的是，还检测到了几种染色质相关蛋白，例如锌指和转录因子，但迄今为止尚不清楚它们是否与精子染色质相关。这提供了有关核蛋白的额外信息，这些核蛋白可能与表观遗传标记、正确受精和胚胎发育相关。
Generating a catalogue of sperm nuclear proteins is an important first step towards the clarification of the function of the paternal chromatin transmitted to the oocyte upon fertilization. With this goal, sperm nuclei were obtained through CTAB treatment and isolated to over 99.9% purity without any tail fragments, acrosome or mitochondria as assessed by optical microscopy and transmission electron microscopy. The nuclear proteins were extracted and separated in 2-D and 1-D gels and the 2-D spots and 1-D bands were excised and analysed to identify the proteins through LC-MS/MS. With this approach, 403 different proteins have been identified from the isolated sperm nuclei. The most abundant family of proteins identified are the histones, for which several novel members had not been reported previously as present in the spermatogenic cell line or in the human mature spermatozoa. More than half (52.6%) of the proteins had not been detected in the previous human whole sperm cell proteome reports. Of relevance, several chromatin-related proteins, such as zinc fingers and transcription factors, so far not known to be associated with the sperm chromatin, have also been detected. This provides additional information about the nuclear proteins that are potentially relevant for epigenetic marking, proper fertilization and embryo development.