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Journal of cell science1997; 110 ( Pt 15); 1821-1829; doi: 10.1242/jcs.110.15.1821

Glyceraldehyde 3-phosphate dehydrogenase is bound to the fibrous sheath of mammalian spermatozoa.

Abstract: Evidence is provided that the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase is covalently linked to the fibrous sheath. The fibrous sheath is a typical structure of mammalian spermatozoa surrounding the axoneme in the principal piece of the flagellum. More than 90% of boar sperm glyceraldehyde 3-phosphate dehydrogenase activity is sedimented after cell disintegration by centrifugation. Detergents, different salt concentrations or short term incubation with chymotrypsin do not solubilize the enzyme, whereas digestion with trypsin or elastase does. Short term incubation with trypsin (15 minutes) even resulted in an activation of glyceraldehyde 3-phosphate dehydrogenase. Purification on phenyl-Sepharose yielded a homogeneous glyceraldehyde 3-phosphate dehydrogenase as judged from gel electrophoresis SDS-PAGE and native gradient PAGE. The molecular masses are 41.5 and 238 kDa, respectively, suggesting native glyceraldehyde 3-phosphate dehydrogenase to be a hexamer. Rabbit polyclonal antibodies raised to purified glyceraldehyde 3-phosphate dehydrogenase show a high specificity for mammalian spermatozoal glyceraldehyde 3-phosphate dehydrogenase, while other proteins of boar spermatozoa or the muscle glyceraldehyde 3-phosphate dehydrogenase are not labelled. Immunogold staining performed in a post-embedding procedure reveals the localization of glyceraldehyde 3-phosphate dehydrogenase along the fibrous sheath in spermatozoa of boar, bull, rat, stallion and man. Other structures such as the cell membrane, dense fibres, the axoneme or the mitochondria are free of label. During the process of sperm maturation, most of the cytoplasm of the sperm midpiece is removed as droplets during the passage through the epididymis. The labelling of this cytoplasm, in immature boar spermatozoa and in the droplets, indicates that glyceraldehyde 3-phosphate dehydrogenase is completely removed from the midpiece during sperm maturation in the epididymis. The inverse compartmentation of the glycolytic enzyme and mitochondria in the mammalian sperm flagella suggests that ATP-production in the principal piece mainly occurs by glycolysis and in the midpiece by respiration.
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Publication Date: 1997-08-01 PubMed ID: 9264469DOI: 10.1242/jcs.110.15.1821Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research investigates the binding of the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase to the fibrous sheath of mammalian spermatozoa, suggesting a key role in energy production during sperm movement.

Research Methods and Findings

  • The researchers started by observing that after disintegration of boar sperm cells by centrifugation, over 90% of the glyceraldehyde 3-phosphate dehydrogenase activity was found in the sediment, indicating that this enzyme is predominantly located in the solid parts of the cell.
  • Attempts to release the enzyme from these parts using detergents, different salt concentrations, or chymotrypsin didn’t succeed significantly. However, treatment with trypsin or elastase was effective, which suggests a very firm binding. Trypsin also activated the enzyme, signaling the possibility of a physiological interaction.
  • Gel electrophoresis methods were used for purification and identification, showing that the enzyme is a hexamer with molecular masses of 41.5 and 238 kDa.
  • Rabbit polyclonal antibodies specifically recognized this exact spermatozoal form of the enzyme, confirming its unique identity.
  • Immunogold staining further confirmed the location of the enzyme along the fibrous sheath in the sperm of different mammals, while it was absent from other sperm and cellular structures.

Implications and Conclusions

  • The researchers noted that the enzyme also appeared in the immature sperm and was then removed during maturation, suggesting a regulatory role of the epididymis in producing mature sperm with optimized structure and function.
  • The specific location of the glyceraldehyde 3-phosphate dehydrogenase within the sperm structure, and the inverse relationship to the mitochondria, suggest it plays a central role in ATP, the primary energy source for cell function, generation within the principal piece of the sperm via glycolysis.
  • The study provided key insights into the importance of the exact position of key enzymes, such as the glyceraldehyde 3-phosphate dehydrogenase, in the functional differentiation of regions within the sperm, and how this may contribute to their motility and fertilizing capabilities.

Cite This Article

APA
Westhoff D, Kamp G. (1997). Glyceraldehyde 3-phosphate dehydrogenase is bound to the fibrous sheath of mammalian spermatozoa. J Cell Sci, 110 ( Pt 15), 1821-1829. https://doi.org/10.1242/jcs.110.15.1821

Publication

Journal of cell science
ISSN: 0021-9533
NlmUniqueID: 0052457
Country: England
Language: English
Volume: 110 ( Pt 15)
Pages: 1821-1829

Researcher Affiliations

Westhoff, D
  • Institut für Zoophysiologie, Westfälische Wilhelms-Universität Münster, Germany.
Kamp, G

    MeSH Terms

    • Animals
    • Antibody Specificity
    • Cattle
    • Cytoplasm / enzymology
    • Electrophoresis, Polyacrylamide Gel
    • Glyceraldehyde-3-Phosphate Dehydrogenases / chemistry
    • Glyceraldehyde-3-Phosphate Dehydrogenases / immunology
    • Glyceraldehyde-3-Phosphate Dehydrogenases / isolation & purification
    • Glyceraldehyde-3-Phosphate Dehydrogenases / metabolism
    • Horses
    • Humans
    • Immunohistochemistry
    • Male
    • Mitochondria / enzymology
    • Molecular Weight
    • Rabbits
    • Rats
    • Solubility
    • Sperm Maturation
    • Sperm Tail / enzymology
    • Sperm Tail / ultrastructure
    • Swine

    Citations

    This article has been cited 43 times.
    1. Balbach M, Ghanem L, Violante S, Kyaw A, Romarowski A, Cross JR, Visconti PE, Levin LR, Buck J. Capacitation induces changes in metabolic pathways supporting motility of epididymal and ejaculated sperm. Front Cell Dev Biol 2023;11:1160154.
      doi: 10.3389/fcell.2023.1160154pubmed: 37440924google scholar: lookup
    2. Zhang S, Wang C, Wang Y, Zhang H, Xu C, Cheng Y, Yuan Y, Sha J, Guo X, Cui Y. A novel protein encoded by circRsrc1 regulates mitochondrial ribosome assembly and translation during spermatogenesis. BMC Biol 2023 Apr 24;21(1):94.
      doi: 10.1186/s12915-023-01597-zpubmed: 37095490google scholar: lookup
    3. Castellini C, D'Andrea S, Cordeschi G, Totaro M, Parisi A, Di Emidio G, Tatone C, Francavilla S, Barbonetti A. Pathophysiology of Mitochondrial Dysfunction in Human Spermatozoa: Focus on Energetic Metabolism, Oxidative Stress and Apoptosis. Antioxidants (Basel) 2021 Apr 28;10(5).
      doi: 10.3390/antiox10050695pubmed: 33924936google scholar: lookup
    4. Gomes FP, Park R, Viana AG, Fernandez-Costa C, Topper E, Kaya A, Memili E, Yates JR 3rd, Moura AA. Protein signatures of seminal plasma from bulls with contrasting frozen-thawed sperm viability. Sci Rep 2020 Sep 4;10(1):14661.
      doi: 10.1038/s41598-020-71015-9pubmed: 32887897google scholar: lookup
    5. Balbach M, Gervasi MG, Hidalgo DM, Visconti PE, Levin LR, Buck J. Metabolic changes in mouse sperm during capacitation†. Biol Reprod 2020 Oct 5;103(4):791-801.
      doi: 10.1093/biolre/ioaa114pubmed: 32614044google scholar: lookup
    6. Vitavska O, Wieczorek H. Putative role of an SLC45 H(+)/sugar cotransporter in mammalian spermatozoa. Pflugers Arch 2017 Nov;469(11):1433-1442.
      doi: 10.1007/s00424-017-2024-9pubmed: 28689241google scholar: lookup
    7. Keighren MA, Flockhart JH, West JD. Survival of glucose phosphate isomerase null somatic cells and germ cells in adult mouse chimaeras. Biol Open 2016 May 15;5(5):596-610.
      doi: 10.1242/bio.017111pubmed: 27103217google scholar: lookup
    8. Paoli D, Pelloni M, Gallo M, Coltrinari G, Lombardo F, Lenzi A, Gandini L. Sperm glyceraldehyde 3-phosphate dehydrogenase gene expression in asthenozoospermic spermatozoa. Asian J Androl 2017 Jul-Aug;19(4):409-413.
      doi: 10.4103/1008-682X.173934pubmed: 27080476google scholar: lookup
    9. Segretain D, Gilleron J, Bacro JN, Di Marco M, Carette D, Pointis G. Ultrastructural localization and distribution of Nardilysin in mammalian male germ cells. Basic Clin Androl 2016;26:5.
      doi: 10.1186/s12610-016-0032-9pubmed: 27051521google scholar: lookup
    10. Danshina PV, Qu W, Temple BR, Rojas RJ, Miley MJ, Machius M, Betts L, O'Brien DA. Structural analyses to identify selective inhibitors of glyceraldehyde 3-phosphate dehydrogenase-S, a sperm-specific glycolytic enzyme. Mol Hum Reprod 2016 Jun;22(6):410-26.
      doi: 10.1093/molehr/gaw016pubmed: 26921398google scholar: lookup
    11. Yin Y, Liu L, Yang C, Lin C, Veith GM, Wang C, Sutovsky P, Zhou P, Ma L. Cell Autonomous and Nonautonomous Function of CUL4B in Mouse Spermatogenesis. J Biol Chem 2016 Mar 25;291(13):6923-35.
      doi: 10.1074/jbc.M115.699660pubmed: 26846852google scholar: lookup
    12. Zhang L, Yang P, Bian X, Zhang Q, Ullah S, Waqas Y, Chen X, Liu Y, Chen W, Le Y, Chen B, Wang S, Chen Q. Modification of sperm morphology during long-term sperm storage in the reproductive tract of the Chinese soft-shelled turtle, Pelodiscus sinensis. Sci Rep 2015 Nov 5;5:16096.
      doi: 10.1038/srep16096pubmed: 26537569google scholar: lookup
    13. Chandramouli KH, Reish D, Zhang H, Qian PY, Ravasi T. Proteomic Changes Associated with Successive Reproductive Periods in Male Polychaetous Neanthes arenaceodentata. Sci Rep 2015 Sep 4;5:13561.
      doi: 10.1038/srep13561pubmed: 26337980google scholar: lookup
    14. du Plessis SS, Agarwal A, Mohanty G, van der Linde M. Oxidative phosphorylation versus glycolysis: what fuel do spermatozoa use?. Asian J Androl 2015 Mar-Apr;17(2):230-5.
      doi: 10.4103/1008-682X.135123pubmed: 25475660google scholar: lookup
    15. Tantibhedhyangkul J, Hawkins KC, Dai Q, Mu K, Dunn CN, Miller SE, Price TM. Expression of a mitochondrial progesterone receptor in human spermatozoa correlates with a progestin-dependent increase in mitochondrial membrane potential. Andrology 2014 Nov;2(6):875-83.
      doi: 10.1111/j.2047-2927.2014.00263.xpubmed: 25187426google scholar: lookup
    16. Mukai C, Gao L, Bergkvist M, Nelson JL, Hinchman MM, Travis AJ. Biomimicry enhances sequential reactions of tethered glycolytic enzymes, TPI and GAPDHS. PLoS One 2013;8(4):e61434.
      doi: 10.1371/journal.pone.0061434pubmed: 23626684google scholar: lookup
    17. Nakamura N, Dai Q, Williams J, Goulding EH, Willis WD, Brown PR, Eddy EM. Disruption of a spermatogenic cell-specific mouse enolase 4 (eno4) gene causes sperm structural defects and male infertility. Biol Reprod 2013 Apr;88(4):90.
      doi: 10.1095/biolreprod.112.107128pubmed: 23446454google scholar: lookup
    18. Amaral A, Castillo J, Estanyol JM, Ballescà JL, Ramalho-Santos J, Oliva R. Human sperm tail proteome suggests new endogenous metabolic pathways. Mol Cell Proteomics 2013 Feb;12(2):330-42.
      doi: 10.1074/mcp.M112.020552pubmed: 23161514google scholar: lookup
    19. Fourie J, Loskutoff N, Huyser C. Treatment of human sperm with serine protease during density gradient centrifugation. J Assist Reprod Genet 2012 Nov;29(11):1273-9.
      doi: 10.1007/s10815-012-9851-6pubmed: 22956335google scholar: lookup
    20. Mukai C, Travis AJ. What sperm can teach us about energy production. Reprod Domest Anim 2012 Aug;47 Suppl 4(0 4):164-9.
      doi: 10.1111/j.1439-0531.2012.02071.xpubmed: 22827366google scholar: lookup
    21. Joice AC, Lyda TL, Sayce AC, Verplaetse E, Morris MT, Michels PA, Robinson DR, Morris JC. Extra-glycosomal localisation of Trypanosoma brucei hexokinase 2. Int J Parasitol 2012 Apr;42(4):401-9.
      doi: 10.1016/j.ijpara.2012.02.008pubmed: 22619756google scholar: lookup
    22. Kuravsky ML, Aleshin VV, Frishman D, Muronetz VI. Testis-specific glyceraldehyde-3-phosphate dehydrogenase: origin and evolution. BMC Evol Biol 2011 Jun 10;11:160.
      doi: 10.1186/1471-2148-11-160pubmed: 21663662google scholar: lookup
    23. Odet F, Gabel SA, Williams J, London RE, Goldberg E, Eddy EM. Lactate dehydrogenase C and energy metabolism in mouse sperm. Biol Reprod 2011 Sep;85(3):556-64.
      doi: 10.1095/biolreprod.111.091546pubmed: 21565994google scholar: lookup
    24. Li YF, He W, Kim YH, Mandal A, Digilio L, Klotz K, Flickinger CJ, Herr JC. CABYR isoforms expressed in late steps of spermiogenesis bind with AKAPs and ropporin in mouse sperm fibrous sheath. Reprod Biol Endocrinol 2010 Aug 23;8:101.
      doi: 10.1186/1477-7827-8-101pubmed: 20731842google scholar: lookup
    25. Terrell KA, Wildt DE, Anthony NM, Bavister BD, Leibo SP, Penfold LM, Marker LL, Crosier AE. Evidence for compromised metabolic function and limited glucose uptake in spermatozoa from the teratospermic domestic cat (Felis catus) and cheetah (Acinonyx jubatus). Biol Reprod 2010 Nov;83(5):833-41.
      doi: 10.1095/biolreprod.110.085639pubmed: 20650882google scholar: lookup
    26. Nakamura N, Mori C, Eddy EM. Molecular complex of three testis-specific isozymes associated with the mouse sperm fibrous sheath: hexokinase 1, phosphofructokinase M, and glutathione S-transferase mu class 5. Biol Reprod 2010 Mar;82(3):504-15.
      doi: 10.1095/biolreprod.109.080580pubmed: 19889946google scholar: lookup
    27. Mukai C, Bergkvist M, Nelson JL, Travis AJ. Sequential reactions of surface- tethered glycolytic enzymes. Chem Biol 2009 Sep 25;16(9):1013-20.
      doi: 10.1016/j.chembiol.2009.08.009pubmed: 19778729google scholar: lookup
    28. Frayne J, Taylor A, Cameron G, Hadfield AT. Structure of insoluble rat sperm glyceraldehyde-3-phosphate dehydrogenase (GAPDH) via heterotetramer formation with Escherichia coli GAPDH reveals target for contraceptive design. J Biol Chem 2009 Aug 21;284(34):22703-12.
      doi: 10.1074/jbc.M109.004648pubmed: 19542219google scholar: lookup
    29. Yan W. Male infertility caused by spermiogenic defects: lessons from gene knockouts. Mol Cell Endocrinol 2009 Jul 10;306(1-2):24-32.
      doi: 10.1016/j.mce.2009.03.003pubmed: 19481682google scholar: lookup
    30. Simpson IA, Dwyer D, Malide D, Moley KH, Travis A, Vannucci SJ. The facilitative glucose transporter GLUT3: 20 years of distinction. Am J Physiol Endocrinol Metab 2008 Aug;295(2):E242-53.
      doi: 10.1152/ajpendo.90388.2008pubmed: 18577699google scholar: lookup
    31. Mitchell DR. The evolution of eukaryotic cilia and flagella as motile and sensory organelles. Adv Exp Med Biol 2007;607:130-40.
      doi: 10.1007/978-0-387-74021-8_11pubmed: 17977465google scholar: lookup
    32. Kim YH, Haidl G, Schaefer M, Egner U, Mandal A, Herr JC. Compartmentalization of a unique ADP/ATP carrier protein SFEC (Sperm Flagellar Energy Carrier, AAC4) with glycolytic enzymes in the fibrous sheath of the human sperm flagellar principal piece. Dev Biol 2007 Feb 15;302(2):463-76.
      doi: 10.1016/j.ydbio.2006.10.004pubmed: 17137571google scholar: lookup
    33. Mitchell BF, Pedersen LB, Feely M, Rosenbaum JL, Mitchell DR. ATP production in Chlamydomonas reinhardtii flagella by glycolytic enzymes. Mol Biol Cell 2005 Oct;16(10):4509-18.
      doi: 10.1091/mbc.e05-04-0347pubmed: 16030251google scholar: lookup
    34. Miki K, Qu W, Goulding EH, Willis WD, Bunch DO, Strader LF, Perreault SD, Eddy EM, O'Brien DA. Glyceraldehyde 3-phosphate dehydrogenase-S, a sperm-specific glycolytic enzyme, is required for sperm motility and male fertility. Proc Natl Acad Sci U S A 2004 Nov 23;101(47):16501-6.
      doi: 10.1073/pnas.0407708101pubmed: 15546993google scholar: lookup
    35. Travis AJ, Foster JA, Rosenbaum NA, Visconti PE, Gerton GL, Kopf GS, Moss SB. Targeting of a germ cell-specific type 1 hexokinase lacking a porin-binding domain to the mitochondria as well as to the head and fibrous sheath of murine spermatozoa. Mol Biol Cell 1998 Feb;9(2):263-76.
      doi: 10.1091/mbc.9.2.263pubmed: 9450953google scholar: lookup
    36. Valente R, Machado AM, Pericuesta E, García-Párraga D, Artilheiro N, Alves F, Sousa-Pinto I, Pinto B, Cordeiro JM, Ruivo R, Gutiérrez-Adán A, Castro LFC. Parallel Erosion of a Testis-Specific Na+/K+ ATPase in Three Mammalian Lineages Sheds Light into the Evolution of Spermatozoa Energetics. Genome Biol Evol 2026 Jan 31;18(2).
      doi: 10.1093/gbe/evaf246pubmed: 41626717google scholar: lookup
    37. Graffeo ML, Dunleavy JEM, Houston BJ, O'Bryan MK. Sperm mitochondrial sheath formation - how and why?. Nat Rev Urol 2025 Nov 11;.
      doi: 10.1038/s41585-025-01109-4pubmed: 41219388google scholar: lookup
    38. Graffeo ML, Nguyen J, Parast FY, Dunleavy JEM, Korneev D, Yang H, Okuda H, O'Connor AE, Conrad DF, Nosrati R, Houston BJ, O'Bryan MK. A novel mechanism of sperm midpiece epididymal maturation and the role of CCDC112 in sperm midpiece formation and establishing an optimal flagella waveform. Cell Commun Signal 2025 Jul 1;23(1):319.
      doi: 10.1186/s12964-025-02320-xpubmed: 40598224google scholar: lookup
    39. Guseva EA, Buev VS, Mirzaeva SE, Pletnev PI, Dontsova OA, Sergiev PV. Structure and Composition of Spermatozoa Fibrous Sheath in Diverse Groups of Metazoa. Int J Mol Sci 2024 Jul 12;25(14).
      doi: 10.3390/ijms25147663pubmed: 39062905google scholar: lookup
    40. Wu H, Zhang Y, Li Y, Sun S, Zhang J, Xie Q, Dong Y, Zhou S, Sha X, Li K, Chen J, Zhang X, Gao Y, Shen Q, Wang G, Zha X, Duan Z, Tang D, Xu C, Geng H, Lv M, Xu Y, Zhou P, Wei Z, Hua R, Cao Y, Liu M, He X. Adenylate kinase phosphate energy shuttle underlies energetic communication in flagellar axonemes. Sci China Life Sci 2024 Aug;67(8):1697-1714.
      doi: 10.1007/s11427-023-2539-1pubmed: 38761355google scholar: lookup
    41. Rosyada ZNA, Pardede BP, Kaiin EM, Gunawan M, Maulana T, Said S, Tumbelaka LITA, Solihin DD, Ulum MF, Purwantara B. A proteomic approach to identifying spermatozoa proteins in Indonesian native Madura bulls. Front Vet Sci 2023;10:1287676.
      doi: 10.3389/fvets.2023.1287676pubmed: 38111731google scholar: lookup
    42. Naletova I, Schmalhausen E, Tomasello B, Pozdyshev D, Attanasio F, Muronetz V. The role of sperm-specific glyceraldehyde-3-phosphate dehydrogenase in the development of pathologies-from asthenozoospermia to carcinogenesis. Front Mol Biosci 2023;10:1256963.
      doi: 10.3389/fmolb.2023.1256963pubmed: 37711387google scholar: lookup
    43. Jarnot P, Ziemska-Legiecka J, Grynberg M, Gruca A. Insights from analyses of low complexity regions with canonical methods for protein sequence comparison. Brief Bioinform 2022 Sep 20;23(5).
      doi: 10.1093/bib/bbac299pubmed: 35914952google scholar: lookup
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