Molecular Pathways Associated with Sperm Biofunction Are Not Affected by the Presence of Round Cell and Leukocyte Proteins in Human Sperm Proteome.
Molecular Pathways Associated with Sperm Biofunction Are Not Affected by the Presence of Round Cell and Leukocyte Proteins in Human Sperm Proteome.
复制标题
与精子生物功能相关的分子途径不受人类精子蛋白质组中圆细胞和白细胞蛋白存在的影响。
DOI：
10.1021/acs.jproteome.8b00829
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发表时间：
2019
影响因子：
4.4
通讯作者：
Samanta,Luna
中科院分区：
文献类型：
--
作者：
PannerSelvam,ManeshKumar;Agarwal,Ashok;Dias,TâniaR;Martins,AnaD;Baskaran,Saradha;Samanta,Luna
在人类精子蛋白质组学实验中，白细胞和圆细胞蛋白可能污染精子蛋白质组，影响生物信息学结果。本研究的主要目的是通过蛋白质组学和生物信息学工具，确定这些蛋白质，特别是来自白细胞的蛋白质在识别精子功能通路中的可能干扰。我们采用液相色谱-串联质谱法对精子蛋白质组进行了评价（1）圆形细胞和白细胞≥1 × 106/mL的精液，（2）圆形细胞和白细胞≥ 1 × 106/mL的精液，（3）圆形细胞<1 × 106/mL的精液，（4）圆形细胞<1 × 106/mL的精液，（5）圆形细胞<1 × 106/mL的精液。（4）圆形细胞<1 × 106/mL的标本经65%密度梯度离心。单纯白细胞培养作为对照组。白细胞≥1 × 106/mL和圆形细胞<1 × 106/mL的精子样本之间的保守DEP（精子和白细胞共有）差异可忽略不计。第1、2、3和4组与对照组之间的比较分析显示，白细胞蛋白的存在不会显著改变精子蛋白质组中所鉴定的典型途径或生物学功能的活化z评分。我们的实验结果表明，圆细胞和白细胞蛋白的存在并不影响与人类精子蛋白功能相关的分子途径的鉴定。因此，使用纯冷冻精液样本进行蛋白质组学研究对下游生物信息学分析无显著影响。
In human sperm proteomic experiments, leukocyte and round cell proteins may contaminate the sperm proteome and affect the bioinformatic results. The main objective of this study was to identify the possible interference of these proteins, especially from leukocytes, in identification of sperm functional pathways through proteomic and bioinformatic tools. We have evaluated the sperm proteome by liquid chromatography–tandem mass spectrometry (LC–MS/MS) in four groups: (1) neat semen with round cells and leukocytes ≥1 × 106/mL; (2) samples with round cells and leukocytes ≥1 × 106/mL processed by 65% density gradient centrifugation; (3) neat semen with round cells <1 × 106/mL; and (4) samples with round cells <1 × 106/mL processed by 65% density gradient centrifugation. Pure leukocyte culture was used as a control group. The difference in the conserved DEPs (common to both sperm and leukocytes) between the sperm samples with leukocytes ≥1 × 106/mL and round cells <1 × 106/mL was negligible. Comparative analysis between groups 1, 2, 3, and 4 with the control group revealed that the presence of leukocyte proteins does not significantly alter the activationz-score of the identified canonical pathways or biological functions in sperm proteome. Our experimental results demonstrate that the presence of round cell and leukocyte proteins do not affect the identification of the molecular pathways associated with human spermatozoa protein function. Hence, the use of neat frozen semen samples for proteomic studies showed no significant impact on the downstream bioinformatic analysis.