Localization and identification of sumoylated proteins in human sperm: excessive sumoylation is a marker of defective spermatozoa
Localization and identification of sumoylated proteins in human sperm: excessive sumoylation is a marker of defective spermatozoa
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DOI：
10.1093/humrep/des317
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发表时间：
2013-01-01
影响因子：
6.1
通讯作者：
Callaway, Myrasol
中科院分区：
文献类型：
--
作者：
Vigodner, Margarita;Shrivastava, Vibha;Callaway, Myrasol
背景：类小泛素化是一种翻译后修饰，参与许多细胞事件的调控。然而，它在人类精子功能中的作用尚未被characterized.METHODS和RESULTS：在这项研究中，免疫荧光和电子显微镜显示，小泛素样修饰物（SUMO）SUMO 1和SUMO 2/3是高度富集在人类精子的颈部区域，这是与多余的核包膜，也可检测到在鞭毛和一些头部区域。在小鼠和果蝇精子中也观察到了类似的SUMO定位模式。与正常精子相比，非运动、双尾、卷曲尾、畸形、小头（小头）和无头（无头）精子在其颈部和尾部区域表现出异常高水平的SUMO化。许多sumo化的蛋白质，范围从20至260 kDa，通过蛋白质印迹法检测，并通过质谱法鉴定，和55 SUMO目标，特别是在人类精子中，而不是在控制部分，对应于鞭毛蛋白，蛋白质参与精子的成熟和分化，热休克蛋白和重要的糖酵解和线粒体酶。确定的靶点包括在生殖细胞和精子中具有特异性功能的蛋白质，如热休克相关的70-kDa蛋白2、外致密纤维蛋白3、A-激酶锚蛋白3和4、L-乳酸脱氢酶C、与X染色体B/F上的细胞核相关的精子蛋白、含缬草酸蛋白的蛋白、明胶、组蛋白H4和泛素。免疫共沉淀实验证实了sumoylation的semenogelin，并表明，一些精子蛋白修饰的sumoylation和ubiquitination同时。结论：许多蛋白质修饰的sumoylation在人类精子;过度sumoylation是有缺陷的精子的标志。
BACKGROUND: Sumoylation is a type of post-translational modification that is implicated in the regulation of numerous cellular events. However, its role in the function of human sperm has not yet been characterized.METHODS AND RESULTS: In this study, both immunofluorescence and electron microscopy revealed that small ubiquitin-like modifiers (SUMO) SUMO1 and SUMO2/3 were highly enriched in the neck area of human sperm that is associated with the redundant nuclear envelope and were also detectable in the flagella and some head regions. Similar localization patterns of SUMO were also observed in mouse and fly sperm. Nonmotile, two-tailed, curled tailed, misshapen, microcephalic (small head) and aciphalic (no head) sperm exhibited abnormally high levels of sumoylation in their neck and tail regions relative to normal sperm. Numerous sumoylated proteins, ranging from 20 to 260 kDa, were detected via western blotting and identified by mass spectrometry, and 55 SUMO targets that were present specifically in human sperm, and not in the control fraction, corresponded to flagella proteins, proteins involved in the maturation and differentiation of sperm, heat shock proteins and important glycolytic and mitochondrial enzymes. The targets that were identified included proteins with specific functions in germ cells and sperm, such as heat shock-related 70-kDa protein 2, outer dense fiber protein 3, A-kinase anchor proteins 3 and 4, L-lactate dehydrogenase C, sperm protein associated with the nucleus on the X chromosome B/F, valosin-containing protein, seminogelins, histone H4 and ubiquitin. Coimmunoprecipitation experiments confirmed the sumoylation of semenogelin and indicated that some sperm proteins are modified by sumoylation and ubiquitination simultaneously.CONCLUSIONS: Numerous proteins are modified by sumoylation in human sperm; excessive sumoylation is a marker of defective spermatozoa.