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Molecular Pathways Associated with Sperm Biofunction Are Not Affected by the Presence of Round Cell and Leukocyte Proteins in Human Sperm Proteome.
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2019-01-11 , DOI: 10.1021/acs.jproteome.8b00829 Manesh Kumar Panner Selvam 1 , Ashok Agarwal 1 , Tânia R Dias 1, 2, 3 , Ana D Martins 1, 3 , Saradha Baskaran 1 , Luna Samanta 1, 4
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2019-01-11 , DOI: 10.1021/acs.jproteome.8b00829 Manesh Kumar Panner Selvam 1 , Ashok Agarwal 1 , Tânia R Dias 1, 2, 3 , Ana D Martins 1, 3 , Saradha Baskaran 1 , Luna Samanta 1, 4
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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 activation z-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.
中文翻译：
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形细胞和白细胞蛋白的影响。
在人类精子蛋白质组学实验中，白细胞和圆形细胞蛋白可能污染精子蛋白质组并影响生物信息学结果。这项研究的主要目的是通过蛋白质组学和生物信息学工具，鉴定这些蛋白质，特别是白细胞的蛋白质，可能对精子功能途径的干扰。我们通过液相色谱-串联质谱法（LC-MS / MS）评估了四组精子蛋白质组：（1）精液中含有圆形细胞和白细胞≥1×106 / mL的纯净精液；（2）65％密度梯度离心处理的圆细胞和白细胞≥1×106 / mL的样品；（3）精液中圆形细胞<1×106 / mL；（4）通过65％密度梯度离心处理的圆形细胞<1×106 / mL的样品。纯白细胞培养用作对照组。白细胞≥1×106 / mL和圆形细胞<1×106 / mL的精子样品之间的保守DEPs（精子和白细胞共同）的差异可以忽略不计。组1、2、3和4与对照组之间的比较分析表明，白细胞蛋白的存在不会显着改变精子蛋白质组中已确定的经典途径或生物学功能的激活z评分。我们的实验结果表明，圆形细胞和白细胞蛋白的存在不影响与人类精子蛋白功能相关的分子途径的鉴定。因此，使用纯净的冷冻精液样品进行蛋白质组学研究对下游生物信息学分析没有显着影响。
更新日期：2019-02-07
中文翻译：
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形细胞和白细胞蛋白的影响。
在人类精子蛋白质组学实验中，白细胞和圆形细胞蛋白可能污染精子蛋白质组并影响生物信息学结果。这项研究的主要目的是通过蛋白质组学和生物信息学工具，鉴定这些蛋白质，特别是白细胞的蛋白质，可能对精子功能途径的干扰。我们通过液相色谱-串联质谱法（LC-MS / MS）评估了四组精子蛋白质组：（1）精液中含有圆形细胞和白细胞≥1×106 / mL的纯净精液；（2）65％密度梯度离心处理的圆细胞和白细胞≥1×106 / mL的样品；（3）精液中圆形细胞<1×106 / mL；（4）通过65％密度梯度离心处理的圆形细胞<1×106 / mL的样品。纯白细胞培养用作对照组。白细胞≥1×106 / mL和圆形细胞<1×106 / mL的精子样品之间的保守DEPs（精子和白细胞共同）的差异可以忽略不计。组1、2、3和4与对照组之间的比较分析表明，白细胞蛋白的存在不会显着改变精子蛋白质组中已确定的经典途径或生物学功能的激活z评分。我们的实验结果表明，圆形细胞和白细胞蛋白的存在不影响与人类精子蛋白功能相关的分子途径的鉴定。因此，使用纯净的冷冻精液样品进行蛋白质组学研究对下游生物信息学分析没有显着影响。
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