Citation Information :
Gunasekaran P, Hemamalini M, Venkatachalam R. Structure Prediction and Binding Site Analysis of Human Sperm Hyaluronidases. Int J Infertil Fetal Med 2022; 13 (3):96-100.
Background: Sperm cell hyaluronidases are important class of enzymes which play a prominent role in hydrolyzing hyaluronan during sperm penetration through the layer of cumulus and zona pellucida of egg. Clinically sperm hyaluronidases play important role in analyzing sperm binding quality during IVF treatments by hyaluronan binding assay. The functions of sperm hyaluronidases such as penetrating the hyaluronan matrix of cumulus layer, inducing hyaluronan-based acrosome reaction were directly related to substrate binding efficiency of these enzymes. Though hyaluronidases were confirmed to play the hyaluronan hydrolyzing role in fertilization, there is no structural evidence to prove their difference in molecular weight-based substrate specificity and their multifunctional activity in fertilization. This work presents the evidence on the binding modes of hyaluronan with sperm cell hyaluronidases using homology modeling and molecular docking methods.
Materials and methods: The different isoforms of human sperm hyaluronidases were modelled using human hyaluronidase 1 (PDB code 2PE4) and the optimized structure of hyaluronan from the crystal structure of bee venom hyaluronidase complexed with HA tetramer (PDB Code: 1vcz) was docked using AUTODOCK TOOLS. The results are evaluated based on the complex binding energy and interaction with substrate binding residues.
Results: The molecular docking results confirm the binding of hyaluronan with HYAL-2 in the deep substrate-binding groove, which was absent in HYAL-3 and HPH-20 hyaluronan complex. The binding of hyaluronan with HPH-20 and HYAL-3, involved in neutral-active domain and residues present in perpendicular beta sheets. The binding mode of HYAL-2 is more on acid-active region of the enzyme which was considered to induce hyaluronan-dependent cell signaling and AR activation.
Conclusion: The results conclude that the substrate accommodation pattern of HYAL-2 was differed from HYAL-3 and HPH-20 where the acid active domain of HYAL-2 may predominantly involve in AR activation and zona pellucida penetration apart from cumulus HA degradation.
Cherr GN, Yudin AI, Overstreet JW. The dual functions of GPI-anchored PH-20: hyaluronidase and intracellular signaling. Matrix Biol 2001;20(8):515–525. DOI:10.1016/s0945-053x(01)00171-8
Modelski MJ, Menlah G, Wang Y, et al. Hyaluronidase 2: a novel germ cell hyaluronidase with epididymal expression and functional roles in mammalian sperm. Biol Reprod 2014;91(5):109. DOI: 10.1095/biolreprod.113.115857
Reese KL, Aravindan RG, Griffiths GS, et al. Acidic hyaluronidase activity is present in mouse sperm and is reduced in the absence of SPAM1: evidence for a role for hyaluronidase 3 in mouse and human sperm. Mol Reprod Dev 2010;77(9):759–772. DOI: 10.1002/mrd.21217
Esterhuizen AD, Franken DR, Bosman E, et al. Relationship between human spermatozoa-hyaluronan-binding assay, conventional semen parameters and fertilisation rates in intracytoplasmic spermatozoa injection. Andrologia 2015;47(7):759–764. DOI: 10.1111/and.12326
Lazarevic J, Wikarczuk M, Somkuti SG, et al. Hyaluronan binding assay (HBA) vs. sperm penetration assay (SPA): Can HBA replace the SPA test in male partner screening before in vitro fertilization? J Exp Clin Assist Reprod 2010;7:2. DOI: 10.1016/j.fertnstert.2007.07.1254
UniProt Consortium. UniProt: a worldwide hub of protein knowledge. Nucleic Acids Res 2019;47(D1):D506–D515. DOI: 10.1093/nar/gky1049
Altschul SF, Gish W, Miller W, et al. Basic local alignment search tool. J Mol Biol 1990;215(3):403–410. DOI: 10.1016/S0022-2836(05)80360-2
Corpet F. Multiple sequence alignment with hierarchical clustering. Nucleic Acids Res 1988;16(22):10881–10890. DOI:10.1093/nar/16.22.10881
Chao KL, Muthukumar L, Herzberg O. Structure of human hyaluronidase-1, a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis. Biochemistry 2007;46(23):6911–6920. DOI: 10.1021/bi700382g
Waterhouse A, Bertoni M, Bienert S, et al. SWISS-MODEL: homology modelling of protein structures and complexes. Nucleic Acids Res 2018;46(W1):W296–W303. DOI: 10.1093/nar/gky427
Bienert S, Waterhouse A, de Beer TAP, et al. The SWISS-MODEL Repository-new features and functionality. Nucleic Acids Res 2017;45(D1):D313–D319, DOI: 10.1093/nar/gkw1132
Guex N, Peitsch MC, Schwede T. Automated comparative protein structure modeling with SWISS-MODEL and Swiss-PdbViewer: a historical perspective. Electrophoresis 2009;30(1):S162–S173. DOI: 10.1002/elps.200900140
Studer G, Rempfer C, Waterhouse AM, et al. QMEANDisCo–distance constraints applied on model quality estimation. Bioinformatics 2020;36(6):1765–1771. DOI: 10.1093/bioinformatics/btz828
Benkert P, Biasini M, Schwede T. Toward the estimation of the absolute quality of individual protein structure models. Bioinformatics 2011;27(3):343–350. DOI:10.1093/bioinformatics/btq662
Bertoni M, Kiefer F, Biasini M, et al. Modeling protein quaternary structure of homo- and hetero-oligomers beyond binary interactions by homology. Sci Rep 2017;7(1):1048010. DOI: 1038/s41598-017-09654-8
Morris GM, Huey R, Lindstrom W, et al. AutoDock4 and AutoDockTools4: automated docking with selective receptor flexibility. J Comput Chem 2009;30(16):2785–2791. DOI: 10.1002/jcc.21256
Marković-Housley Z, Miglierini G, Soldatova L, et al. Crystal structure of hyaluronidase, a major allergen of bee venom. Structure 2000;8(10):1025–1035. DOI: 10.1016/s0969-2126(00)00511-6
DeLano, W. L. (2009). The PyMOL Molecular Graphics System; DeLano Scientific: San Carlos, CA, 2002.
Martin-Deleon PA. Germ-cell hyaluronidases: their roles in sperm function. Int J Androl 2011;34(5 Pt 2):e306–e318. DOI: 10.1111/j.1365-2605.2010.01138.x
Lin Y, Mahan K, Lathrop WF, et al. A hyaluronidase activity of the sperm plasma membrane protein PH-20 enables sperm to penetrate the cumulus cell layer surrounding the egg. J Cell Biol 1994;125(5):1157–1163. DOI: 10.1083/jcb.125.5.1157
Meyer MF, Kreil G, Aschauer H. The soluble hyaluronidase from bull testes is a fragment of the membrane-bound PH-20 enzyme. FEBS Lett 1997;413(2):385–388. DOI: 10.1016/s0014-5793(97)00936-8
Botzki A, Rigden DJ, Braun S, et al. L-Ascorbic acid 6-hexadecanoate, a potent hyaluronidase inhibitor. X-ray structure and molecular modeling of enzyme-inhibitor complexes. J Biol Chem 2004;279(44):45990–45997. DOI:10.1074/jbc.M406146200
Zeng HJ, Yang R, You J, et al. Spectroscopic and docking studies on the binding of liquiritigenin with hyaluronidase for antiallergic mechanism. Scientifica (Cairo) 2016;2016:9178097. DOI: 10.1155/2016/9178097
Hunnicutt GR, Primakoff P, Myles DG. Sperm surface protein PH-20 is bifunctional: one activity is a hyaluronidase and a second, distinct activity is required in secondary sperm-zona binding. Biol Reprod 1996;55(1):80–86. DOI:10.1095/biolreprod55.1.80
Hunnicutt GR, Mahan K, Lathrop WF, et al. Structural relationship of sperm soluble hyaluronidase to the sperm membrane protein PH-20. Biol Reprod 1996;54(6):1343–1349. DOI:10.1095/biolreprod54.6.1343
Stern R, Jedrzejas MJ. Hyaluronidases: their genomics, structures, and mechanisms of action. Chem Rev 2006;106(3):818–839. DOI: 10.1021/cr050247k
