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Proteomic Landscape of Human Spermatozoa: Optimized Extraction Method and Application

Human spermatozoa proteomics exposed to some physical, biological or chemical stressors is being explored. However, there is a lack of optimized sample preparation methods to achieve in-depth protein coverage for sperm cells. Meanwhile, it is not clear whether antibiotics can regulate proteins to af...

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Autores principales: Luo, Mengqi, Su, Tao, Wang, Shisheng, Chen, Jianhai, Lin, Tianhai, Cheng, Qingyuan, Chen, Younan, Gong, Meng, Yang, Hao, Li, Fuping, Zhang, Yong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9776871/
https://www.ncbi.nlm.nih.gov/pubmed/36552826
http://dx.doi.org/10.3390/cells11244064
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author Luo, Mengqi
Su, Tao
Wang, Shisheng
Chen, Jianhai
Lin, Tianhai
Cheng, Qingyuan
Chen, Younan
Gong, Meng
Yang, Hao
Li, Fuping
Zhang, Yong
author_facet Luo, Mengqi
Su, Tao
Wang, Shisheng
Chen, Jianhai
Lin, Tianhai
Cheng, Qingyuan
Chen, Younan
Gong, Meng
Yang, Hao
Li, Fuping
Zhang, Yong
author_sort Luo, Mengqi
collection PubMed
description Human spermatozoa proteomics exposed to some physical, biological or chemical stressors is being explored. However, there is a lack of optimized sample preparation methods to achieve in-depth protein coverage for sperm cells. Meanwhile, it is not clear whether antibiotics can regulate proteins to affect sperm quality. Here, we systematically compared a total of six different protein extraction methods based the combination of three commonly used lysis buffers and physical lysis strategies. The urea buffer combined with ultrasonication (UA-ultrasonication) produced the highest protein extraction rate, leading to the deepest coverage of human sperm proteome (5685 protein groups) from healthy human sperm samples. Since the antibiotics, amoxicillin and clarithromycin, have been widely used against H. pylori infection, we conduct a longitudinal study of sperm proteome via data-independent acquisition tandem mass spectrometry (DIA-MS/MS) on an infected patient during on and off therapy with these two drugs. The semen examination and morphological analysis were performed combined with proteomics analysis. Our results indicated that antibiotics may cause an increase in the sperm concentration and the rate of malformed sperm and disrupt proteome expression in sperm. This work provides an optimized extraction method to characterize the in-depth human sperm proteome and to extend its clinical applications.
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spelling pubmed-97768712022-12-23 Proteomic Landscape of Human Spermatozoa: Optimized Extraction Method and Application Luo, Mengqi Su, Tao Wang, Shisheng Chen, Jianhai Lin, Tianhai Cheng, Qingyuan Chen, Younan Gong, Meng Yang, Hao Li, Fuping Zhang, Yong Cells Article Human spermatozoa proteomics exposed to some physical, biological or chemical stressors is being explored. However, there is a lack of optimized sample preparation methods to achieve in-depth protein coverage for sperm cells. Meanwhile, it is not clear whether antibiotics can regulate proteins to affect sperm quality. Here, we systematically compared a total of six different protein extraction methods based the combination of three commonly used lysis buffers and physical lysis strategies. The urea buffer combined with ultrasonication (UA-ultrasonication) produced the highest protein extraction rate, leading to the deepest coverage of human sperm proteome (5685 protein groups) from healthy human sperm samples. Since the antibiotics, amoxicillin and clarithromycin, have been widely used against H. pylori infection, we conduct a longitudinal study of sperm proteome via data-independent acquisition tandem mass spectrometry (DIA-MS/MS) on an infected patient during on and off therapy with these two drugs. The semen examination and morphological analysis were performed combined with proteomics analysis. Our results indicated that antibiotics may cause an increase in the sperm concentration and the rate of malformed sperm and disrupt proteome expression in sperm. This work provides an optimized extraction method to characterize the in-depth human sperm proteome and to extend its clinical applications. MDPI 2022-12-15 /pmc/articles/PMC9776871/ /pubmed/36552826 http://dx.doi.org/10.3390/cells11244064 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Luo, Mengqi
Su, Tao
Wang, Shisheng
Chen, Jianhai
Lin, Tianhai
Cheng, Qingyuan
Chen, Younan
Gong, Meng
Yang, Hao
Li, Fuping
Zhang, Yong
Proteomic Landscape of Human Spermatozoa: Optimized Extraction Method and Application
title Proteomic Landscape of Human Spermatozoa: Optimized Extraction Method and Application
title_full Proteomic Landscape of Human Spermatozoa: Optimized Extraction Method and Application
title_fullStr Proteomic Landscape of Human Spermatozoa: Optimized Extraction Method and Application
title_full_unstemmed Proteomic Landscape of Human Spermatozoa: Optimized Extraction Method and Application
title_short Proteomic Landscape of Human Spermatozoa: Optimized Extraction Method and Application
title_sort proteomic landscape of human spermatozoa: optimized extraction method and application
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9776871/
https://www.ncbi.nlm.nih.gov/pubmed/36552826
http://dx.doi.org/10.3390/cells11244064
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