Phosphoproteomics and Bioinformatics Analyses Reveal Key Roles of GSK-3 and AKAP4 in Mouse Sperm Capacitation
Protein phosphorylation can induce signal transduction to change sperm motility patterns during sperm capacitation. However, changes in the phosphorylation of sperm proteins in mice are still incompletely understood. Here, capacitation-related phosphorylation in mouse sperms were firstly investigate...
| Main Authors: | , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-10-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/21/19/7283 |
| _version_ | 1797551978643980288 |
|---|---|
| author | Nailis Syifa Jhih-Tian Yang Chang-Shiann Wu Miao-Hsia Lin Wan-Ling Wu Cheng-Wei Lai Sheng-Hsuan Ku Suh-Yuen Liang Yu-Chun Hung Chia-Te Chou Chien-Sheng Wang Yasushi Ishihama Jiahn-Haur Liao Shih-Hsiung Wu Tzu-Hua Wu |
| author_facet | Nailis Syifa Jhih-Tian Yang Chang-Shiann Wu Miao-Hsia Lin Wan-Ling Wu Cheng-Wei Lai Sheng-Hsuan Ku Suh-Yuen Liang Yu-Chun Hung Chia-Te Chou Chien-Sheng Wang Yasushi Ishihama Jiahn-Haur Liao Shih-Hsiung Wu Tzu-Hua Wu |
| author_sort | Nailis Syifa |
| collection | DOAJ |
| description | Protein phosphorylation can induce signal transduction to change sperm motility patterns during sperm capacitation. However, changes in the phosphorylation of sperm proteins in mice are still incompletely understood. Here, capacitation-related phosphorylation in mouse sperms were firstly investigated by label-free quantitative (LFQ) phosphoproteomics coupled with bioinformatics analysis using ingenuity pathway analysis (IPA) methods such as canonical pathway, upstream regulator, and network analysis. Among 1632 phosphopeptides identified at serine, threonine, and tyrosine residues, 1050 novel phosphosites, corresponding to 402 proteins, were reported. Gene heatmaps for IPA canonical pathways showed a novel role for GSK-3 in GP6 signaling pathways associated with capacitation for 60 min. At the same time, the reduction of the abundant isoform-specific GSK-3α expression was shown by western blot (WB) while the LFQ pY of this isoform slightly decreased and then increased. The combined results from WB and LFQ methods explain the less inhibitory phosphorylation of GSK-3α during capacitation and also support the predicted increases in its activity. In addition, pAKAP4 increased at the Y156 site but decreased at the Y811 site in a capacitated state, even though IPA network analysis and WB analysis for overall pAKAP revealed upregulated trends. The potential roles of GSK-3 and AKAP4 in fertility are discussed. |
| first_indexed | 2024-03-10T15:52:22Z |
| format | Article |
| id | doaj.art-072d80db5ca54462a4e58e0f1daf69a4 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T15:52:22Z |
| publishDate | 2020-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-072d80db5ca54462a4e58e0f1daf69a42023-11-20T15:52:13ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-10-012119728310.3390/ijms21197283Phosphoproteomics and Bioinformatics Analyses Reveal Key Roles of GSK-3 and AKAP4 in Mouse Sperm CapacitationNailis Syifa0Jhih-Tian Yang1Chang-Shiann Wu2Miao-Hsia Lin3Wan-Ling Wu4Cheng-Wei Lai5Sheng-Hsuan Ku6Suh-Yuen Liang7Yu-Chun Hung8Chia-Te Chou9Chien-Sheng Wang10Yasushi Ishihama11Jiahn-Haur Liao12Shih-Hsiung Wu13Tzu-Hua Wu14Division of Clinical Pharmacy, School of Pharmacy, College of Pharmacy, Taipei Medical University, Taipei 110, TaiwanPh.D. Program in Microbial Genomics, National Chung Hsing University and Academia Sinica, Taichung 40227, TaiwanDepartment of Information Management, National Formosa University, Yunlin County 632, TaiwanGraduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, JapanInstitute of Biological Chemistry, Academia Sinica, Taipei 115, TaiwanDivision of Clinical Pharmacy, School of Pharmacy, College of Pharmacy, Taipei Medical University, Taipei 110, TaiwanDivision of Clinical Pharmacy, School of Pharmacy, College of Pharmacy, Taipei Medical University, Taipei 110, TaiwanInstitute of Biological Chemistry, Academia Sinica, Taipei 115, TaiwanDivision of Clinical Pharmacy, School of Pharmacy, College of Pharmacy, Taipei Medical University, Taipei 110, TaiwanDivision of Clinical Pharmacy, School of Pharmacy, College of Pharmacy, Taipei Medical University, Taipei 110, TaiwanDivision of Clinical Pharmacy, School of Pharmacy, College of Pharmacy, Taipei Medical University, Taipei 110, TaiwanGraduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, JapanInstitute of Biological Chemistry, Academia Sinica, Taipei 115, TaiwanInstitute of Biological Chemistry, Academia Sinica, Taipei 115, TaiwanDivision of Clinical Pharmacy, School of Pharmacy, College of Pharmacy, Taipei Medical University, Taipei 110, TaiwanProtein phosphorylation can induce signal transduction to change sperm motility patterns during sperm capacitation. However, changes in the phosphorylation of sperm proteins in mice are still incompletely understood. Here, capacitation-related phosphorylation in mouse sperms were firstly investigated by label-free quantitative (LFQ) phosphoproteomics coupled with bioinformatics analysis using ingenuity pathway analysis (IPA) methods such as canonical pathway, upstream regulator, and network analysis. Among 1632 phosphopeptides identified at serine, threonine, and tyrosine residues, 1050 novel phosphosites, corresponding to 402 proteins, were reported. Gene heatmaps for IPA canonical pathways showed a novel role for GSK-3 in GP6 signaling pathways associated with capacitation for 60 min. At the same time, the reduction of the abundant isoform-specific GSK-3α expression was shown by western blot (WB) while the LFQ pY of this isoform slightly decreased and then increased. The combined results from WB and LFQ methods explain the less inhibitory phosphorylation of GSK-3α during capacitation and also support the predicted increases in its activity. In addition, pAKAP4 increased at the Y156 site but decreased at the Y811 site in a capacitated state, even though IPA network analysis and WB analysis for overall pAKAP revealed upregulated trends. The potential roles of GSK-3 and AKAP4 in fertility are discussed.https://www.mdpi.com/1422-0067/21/19/7283spermcapacitationbioinformaticsmouseGSK-3AKAP4 |
| spellingShingle | Nailis Syifa Jhih-Tian Yang Chang-Shiann Wu Miao-Hsia Lin Wan-Ling Wu Cheng-Wei Lai Sheng-Hsuan Ku Suh-Yuen Liang Yu-Chun Hung Chia-Te Chou Chien-Sheng Wang Yasushi Ishihama Jiahn-Haur Liao Shih-Hsiung Wu Tzu-Hua Wu Phosphoproteomics and Bioinformatics Analyses Reveal Key Roles of GSK-3 and AKAP4 in Mouse Sperm Capacitation International Journal of Molecular Sciences sperm capacitation bioinformatics mouse GSK-3 AKAP4 |
| title | Phosphoproteomics and Bioinformatics Analyses Reveal Key Roles of GSK-3 and AKAP4 in Mouse Sperm Capacitation |
| title_full | Phosphoproteomics and Bioinformatics Analyses Reveal Key Roles of GSK-3 and AKAP4 in Mouse Sperm Capacitation |
| title_fullStr | Phosphoproteomics and Bioinformatics Analyses Reveal Key Roles of GSK-3 and AKAP4 in Mouse Sperm Capacitation |
| title_full_unstemmed | Phosphoproteomics and Bioinformatics Analyses Reveal Key Roles of GSK-3 and AKAP4 in Mouse Sperm Capacitation |
| title_short | Phosphoproteomics and Bioinformatics Analyses Reveal Key Roles of GSK-3 and AKAP4 in Mouse Sperm Capacitation |
| title_sort | phosphoproteomics and bioinformatics analyses reveal key roles of gsk 3 and akap4 in mouse sperm capacitation |
| topic | sperm capacitation bioinformatics mouse GSK-3 AKAP4 |
| url | https://www.mdpi.com/1422-0067/21/19/7283 |
| work_keys_str_mv | AT nailissyifa phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation AT jhihtianyang phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation AT changshiannwu phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation AT miaohsialin phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation AT wanlingwu phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation AT chengweilai phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation AT shenghsuanku phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation AT suhyuenliang phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation AT yuchunhung phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation AT chiatechou phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation AT chienshengwang phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation AT yasushiishihama phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation AT jiahnhaurliao phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation AT shihhsiungwu phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation AT tzuhuawu phosphoproteomicsandbioinformaticsanalysesrevealkeyrolesofgsk3andakap4inmousespermcapacitation |