Abstract
The complexities of energy transfer mechanisms in the flagella of mammalian sperm flagella have been intensively investigated and demonstrate significant diversity across species. Enzymatic shuttles, particularly adenylate kinase (AK) and creatine kinase (CK), are pivotal in the efficient transfer of intracellular ATP, showing distinct tissue- and species-specificity. Here, the expression profiles of AK and CK were investigated in mice and found to fall into four subgroups, of which Subgroup III AKs were observed to be unique to the male reproductive system and conserved across chordates. Both AK8 and AK9 were found to be indispensable to male reproduction after analysis of an infertile male cohort. Knockout mouse models showed that AK8 and AK9 were central to promoting sperm motility. Immunoprecipitation combined with mass spectrometry revealed that AK8 and AK9 interact with the radial spoke (RS) of the axoneme. Examination of various human and mouse sperm samples with substructural damage, including the presence of multiple RS subunits, showed that the head of radial spoke 3 acts as an adapter for AK9 in the flagellar axoneme. Using an ATP probe together with metabolomic analysis, it was found that AK8 and AK9 cooperatively regulated ATP transfer in the axoneme, and were concentrated at sites associated with energy consumption in the flagellum. These findings indicate a novel function for RS beyond its structural role, namely, the regulation of ATP transfer. In conclusion, the results expand the functional spectrum of AK proteins and suggest a fresh model regarding ATP transfer within mammalian flagella.
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All the data supporting the results of the present study are provided to the corresponding authors upon request.
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Acknowledgement
This work was supported by National Key Research and Development Program of China (2022YFC2702702, 2021YFC2700901), the National Natural Science Foundation of China (81971441, 82171607, 32000584), the University Outstanding Young Talents Support Program (gxyq2021174), Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences (2019PT310002), Anhui Provincial Natural Science Foundation (2208085Y31), and the Natural Science Foundation of Jiangsu Province (BK20230004). We sincerely appreciate Tao Zhou from Nanjing Wushangzhigao Biomedical Technology Co., LTD and Yangyang Wu from Nanjing Maternity and Child Health Care Hospital for the data analysis of metabolomics; Danhong Qiu for the drawing of the model; Zibin Wang from Nanjing Medical University and Haijian Cai from the Anhui Medical University for the sample preparation and SEM/TEM analysis.
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Wu, H., Zhang, Y., Li, Y. et al. Adenylate kinase phosphate energy shuttle underlies energetic communication in flagellar axonemes. Sci. China Life Sci. (2024). https://doi.org/10.1007/s11427-023-2539-1
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DOI: https://doi.org/10.1007/s11427-023-2539-1