{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,27]],"date-time":"2026-05-27T15:48:40Z","timestamp":1779896920627,"version":"3.53.1"},"reference-count":58,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,7,20]],"date-time":"2021-07-20T00:00:00Z","timestamp":1626739200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell Dev. Biol."],"abstract":"<jats:p>Capacitated spermatozoa initiate fertilization by binding to the zona pellucida (ZP). Defective spermatozoa-ZP binding causes infertility. The sialyl-Lewis(x) (SLeX) sequence is the most abundant terminal sequence on the glycans of human ZP glycoproteins involving in spermatozoa-ZP binding. This study aimed to identify and characterize the SLeX-binding proteins on human spermatozoa. By using affinity chromatography followed by mass spectrometric analysis, chromosome 1 open reading frame 56 (C1orf56) was identified to be a SLeX-binding protein of capacitated spermatozoa. The acrosomal region of spermatozoa possessed C1orf56 immunoreactive signals with intensities that increased after capacitation indicating translocation of C1orf56 to the cell surface during capacitation. Treatment with antibody against C1orf56 inhibited spermatozoa-ZP binding and ZP-induced acrosome reaction. Purified C1orf56 from capacitated spermatozoa bound to human ZP. A pilot clinical study was conducted and found no association between the percentage of capacitated spermatozoa with C1orf56 expression and <jats:italic>in vitro<\/jats:italic> fertilization (IVF) rate in assisted reproduction treatment. However, the percentage of C1orf56 positive spermatozoa in the acrosome-reacted population was significantly (<jats:italic>P<\/jats:italic> &amp;lt; 0.05) lower in cycles with a fertilization rate &amp;lt; 60% when compared to those with a higher fertilization rate, suggesting that C1orf56 may have functions after ZP-binding and acrosome reaction. A larger clinical trial is needed to determine the possible use of sperm C1orf56 content for the prediction of fertilization potential of sperm samples.<\/jats:p>","DOI":"10.3389\/fcell.2021.700396","type":"journal-article","created":{"date-parts":[[2021,7,20]],"date-time":"2021-07-20T10:15:41Z","timestamp":1626776141000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":9,"title":["Identification of Sialyl-Lewis(x)-Interacting Protein on Human Spermatozoa"],"prefix":"10.3389","volume":"9","author":[{"given":"Ying","family":"Wang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weie","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Si","family":"Mei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Panyu","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tsz-Ying","family":"Leung","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cheuk-Lun","family":"Lee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William S. B.","family":"Yeung","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian-Ping","family":"Ou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyan","family":"Liang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip C. N.","family":"Chiu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,7,20]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"280","DOI":"10.1002\/(SICI)1097-0177(200006)218:2<280::AID-DVDY4<3.0.CO;2-0","article-title":"The thrombospondin type 1 repeat (TSR) superfamily: diverse proteins with related roles in neuronal development.","volume":"218","author":"Adams","year":"2000","journal-title":"Dev. Dyn."},{"key":"B2","doi-asserted-by":"publisher","first-page":"1298","DOI":"10.1093\/humrep\/deh798","article-title":"Identification and evaluation of a novel sperm protamine abnormality in a population of infertile males.","volume":"20","author":"Aoki","year":"2005","journal-title":"Hum. Reprod."},{"key":"B3","doi-asserted-by":"publisher","first-page":"3645","DOI":"10.1242\/jcs.01214","article-title":"Tyrosine phosphorylation activates surface chaperones facilitating sperm-zona recognition.","author":"Asquith","year":"2004","journal-title":"J. Cell Sci."},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.aad9946","article-title":"ZP2 peptide beads select human sperm in vitro, decoy mouse sperm in vivo, and provide reversible contraception.","volume":"8","author":"Avella","year":"2016","journal-title":"Sci. Transl. Med."},{"key":"B5","doi-asserted-by":"publisher","first-page":"801","DOI":"10.1083\/jcb.201404025","article-title":"A single domain of the ZP2 zona pellucida protein mediates gamete recognition in mice and humans.","volume":"205","author":"Avella","year":"2014","journal-title":"J. Cell Biol."},{"key":"B6","doi-asserted-by":"publisher","first-page":"279","DOI":"10.1093\/molehr\/gat004","article-title":"The molecular basis of gamete recognition in mice and humans.","volume":"19","author":"Avella","year":"2013","journal-title":"Mol. Hum. Reprod."},{"key":"B7","doi-asserted-by":"publisher","first-page":"897","DOI":"10.1083\/jcb.201203062","article-title":"Human sperm bind to the N-terminal domain of ZP2 in humanized zonae pellucidae in transgenic mice.","volume":"197","author":"Baibakov","year":"2012","journal-title":"J. Cell Biol."},{"key":"B8","doi-asserted-by":"publisher","first-page":"2075","DOI":"10.1016\/s0140-6736(00)05179-5","article-title":"Conventional in-vitro fertilisation versus intracytoplasmic sperm injection for the treatment of non-male-factor infertility: a randomised controlled trial.","volume":"357","author":"Bhattacharya","year":"2001","journal-title":"Lancet"},{"key":"B9","doi-asserted-by":"publisher","first-page":"7970","DOI":"10.1002\/chem.201800451","article-title":"Chemoenzymatic synthesis of asymmetrical multi-antennary N-glycans to dissect glycan-mediated interactions between human sperm and oocytes.","volume":"24","author":"Chinoy","year":"2018","journal-title":"Chemistry"},{"key":"B10","doi-asserted-by":"publisher","first-page":"5378","DOI":"10.1074\/jbc.M607482200","article-title":"Cumulus oophorus-associated glycodelin-C displaces sperm-bound glycodelin-A and -F and stimulates spermatozoa-zona pellucida binding.","volume":"282","author":"Chiu","year":"","journal-title":"J. Biol. Chem."},{"key":"B11","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1242\/jcs.03258","article-title":"Glycodelin-A interacts with fucosyltransferase on human sperm plasma membrane to inhibit spermatozoa-zona pellucida binding.","author":"Chiu","year":"","journal-title":"J. Cell Sci."},{"key":"B12","doi-asserted-by":"publisher","first-page":"25580","DOI":"10.1074\/jbc.M504103200","article-title":"Glycodelin-S in human seminal plasma reduces cholesterol efflux and inhibits capacitation of spermatozoa.","volume":"280","author":"Chiu","year":"2005","journal-title":"J. Biol. Chem."},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1002\/mrd.21398","article-title":"Adrenomedullin enhances the progressive motility and zona pellucida-binding capacity of spermatozoa from normozoospermic but not asthenozoospermic men.","volume":"79","author":"Chiu","year":"2012","journal-title":"Mol. Reprod. Dev."},{"key":"B14","doi-asserted-by":"publisher","first-page":"86","DOI":"10.1016\/j.semcdb.2014.04.016","article-title":"The identity of zona pellucida receptor on spermatozoa: an unresolved issue in developmental biology.","volume":"30","author":"Chiu","year":"2014","journal-title":"Semin. Cell Dev. Biol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"869","DOI":"10.1095\/biolreprod.108.069344","article-title":"Effects of native human zona pellucida glycoproteins 3 and 4 on acrosome reaction and zona pellucida binding of human spermatozoa.","volume":"79","author":"Chiu","year":"","journal-title":"Biol. Reprod."},{"key":"B16","doi-asserted-by":"publisher","first-page":"2721","DOI":"10.1093\/humrep\/deq243","article-title":"Zona pellucida-induced acrosome reaction in human spermatozoa is potentiated by glycodelin-A via down-regulation of extracellular signal-regulated kinases and up-regulation of zona pellucida-induced calcium influx.","volume":"25","author":"Chiu","year":"2010","journal-title":"Hum. Reprod."},{"key":"B17","doi-asserted-by":"publisher","first-page":"1385","DOI":"10.1093\/humrep\/den047","article-title":"Native human zona pellucida glycoproteins: purification and binding properties.","volume":"23","author":"Chiu","year":"","journal-title":"Hum. Reprod."},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1016\/j.fsigen.2020.102313","article-title":"A confirmatory test for sperm in sexual assault samples using a microfluidic-integrated cell phone imaging system.","volume":"48","author":"Deshmukh","year":"2020","journal-title":"Forensic. Sci. Int. Genet."},{"key":"B19","doi-asserted-by":"publisher","first-page":"1136","DOI":"10.1095\/biolreprod57.5.1136","article-title":"Biochemical characterization of sperm agglutination antigen-1, a human sperm surface antigen implicated in gamete interactions.","volume":"57","author":"Diekman","year":"1997","journal-title":"Biol. Reprod."},{"key":"B20","doi-asserted-by":"publisher","first-page":"197","DOI":"10.1002\/mrd.21124","article-title":"The \u201cswitching on\u201d of mammalian spermatozoa: molecular events involved in promotion and regulation of capacitation.","volume":"77","author":"Fraser","year":"2010","journal-title":"Mol. Reprod. Dev."},{"key":"B21","doi-asserted-by":"publisher","first-page":"473","DOI":"10.1387\/ijdb.082583bg","article-title":"Sperm head membrane reorganisation during capacitation.","volume":"52","author":"Gadella","year":"2008","journal-title":"Int. J. Dev. Biol."},{"key":"B22","doi-asserted-by":"publisher","first-page":"379","DOI":"10.1016\/bs.ctdb.2018.01.001","article-title":"The human egg\u2019s zona pellucida.","volume":"130","author":"Gupta","year":"2018","journal-title":"Curr. Top. Dev. Biol."},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.3389\/fcell.2021.619868","article-title":"Human zona pellucida glycoproteins: binding characteristics with human spermatozoa and induction of acrosome reaction.","volume":"9","author":"Gupta","year":"2021","journal-title":"Front. Cell Dev. Biol."},{"key":"B24","doi-asserted-by":"publisher","first-page":"735","DOI":"10.1095\/biolreprod66.3.735","article-title":"SAMP32, a testis-specific, isoantigenic sperm acrosomal membrane-associated protein.","volume":"66","author":"Hao","year":"2002","journal-title":"Biol. Reprod."},{"key":"B25","doi-asserted-by":"publisher","first-page":"996","DOI":"10.1002\/bies.10172","article-title":"Reciprocal raft-receptor interactions and the assembly of adhesion complexes.","volume":"24","author":"Harris","year":"2002","journal-title":"Bioessays"},{"key":"B26","doi-asserted-by":"publisher","first-page":"1444","DOI":"10.1016\/j.fertnstert.2012.11.056","article-title":"Cell membrane proteins from oviductal epithelial cell line protect human spermatozoa from oxidative damage.","volume":"99","author":"Huang","year":"2013","journal-title":"Fertil Steril"},{"key":"B27","doi-asserted-by":"publisher","first-page":"925","DOI":"10.1095\/biolreprod63.3.925","article-title":"Putative creatine kinase M-isoform in human sperm is identifiedas the 70-kilodalton heat shock protein HspA2.","volume":"63","author":"Huszar","year":"2000","journal-title":"Biol. Reprod."},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1002\/advs.201800121","article-title":"A novel on-chip method for differential extraction of sperm in forensic cases.","volume":"5","author":"Inci","year":"2018","journal-title":"Adv. Sci. (Weinh)"},{"key":"B29","doi-asserted-by":"publisher","first-page":"289","DOI":"10.1016\/0141-8130(80)90047-1","article-title":"Assessment of the use of cyanogen bromide-activated Sepharose in analytic and preparative immunoadsorption.","volume":"2","author":"Kennedy","year":"1980","journal-title":"Int. J. Biol. Macromol"},{"key":"B30","doi-asserted-by":"publisher","first-page":"139","DOI":"10.1016\/j.jtbi.2011.11.002","article-title":"The relative contributions of propulsive forces and receptor-ligand binding forces during early contact between spermatozoa and zona pellucida of oocytes.","volume":"294","author":"Kozlovsky","year":"2012","journal-title":"J. Theor. Biol."},{"key":"B31","doi-asserted-by":"publisher","first-page":"381","DOI":"10.1083\/jcb.200811059","article-title":"Lattices, rafts, and scaffolds: domain regulation of receptor signaling at the plasma membrane.","volume":"185","author":"Lajoie","year":"2009","journal-title":"J. Cell Biol."},{"key":"B32","doi-asserted-by":"publisher","first-page":"909","DOI":"10.2337\/db10-1186","article-title":"Glycosylation failure extends to glycoproteins in gestational diabetes mellitus: evidence from reduced alpha2-6 sialylation and impaired immunomodulatory activities of pregnancy-related glycodelin-A.","volume":"60","author":"Lee","year":"2011","journal-title":"Diabetes"},{"key":"B33","doi-asserted-by":"publisher","first-page":"6794","DOI":"10.1128\/MCB.01029-07","article-title":"Loss of zona pellucida binding proteins in the acrosomal matrix disrupts acrosome biogenesis and sperm morphogenesis.","volume":"27","author":"Lin","year":"2007","journal-title":"Mol. Cell Biol."},{"key":"B34","doi-asserted-by":"publisher","first-page":"702","DOI":"10.1093\/humrep\/15.3.702","article-title":"Defective sperm-zona pellucida interaction: a major cause of failure of fertilization in clinical in-vitro fertilization.","volume":"15","author":"Liu","year":"2000","journal-title":"Hum. Reprod."},{"key":"B35","doi-asserted-by":"publisher","first-page":"228","DOI":"10.1093\/humrep\/deh067","article-title":"High frequency of defective sperm-zona pellucida interaction in oligozoospermic infertile men.","volume":"19","author":"Liu","year":"2004","journal-title":"Hum. Reprod."},{"key":"B36","doi-asserted-by":"publisher","first-page":"458","DOI":"10.1093\/molehr\/gaz030","article-title":"The role of galectin-3 in spermatozoa-zona pellucida binding and its association with fertilization in vitro.","volume":"25","author":"Mei","year":"2019","journal-title":"Mol. Hum. Reprod."},{"key":"B37","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1111\/j.1439-0272.1994.tb00774.x","article-title":"Sperm-oocyte interaction: studies on the kinetics of zona pellucida binding and acrosome reaction of human spermatozoa.","volume":"26","author":"Morales","year":"1994","journal-title":"Andrologia"},{"key":"B38","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1016\/j.jri.2009.06.258","article-title":"The biological significance of detergent-resistant membranes in spermatozoa.","volume":"83","author":"Nixon","year":"2009","journal-title":"J. Reprod. Immunol."},{"key":"B39","doi-asserted-by":"publisher","first-page":"122","DOI":"10.1002\/jcp.21575","article-title":"Composition and significance of detergent resistant membranes in mouse spermatozoa.","volume":"218","author":"Nixon","year":"2009","journal-title":"J. Cell Physiol."},{"key":"B40","doi-asserted-by":"publisher","first-page":"568","DOI":"10.4103\/1008-682X.151395","article-title":"The role of the molecular chaperone heat shock protein A2 (HSPA2) in regulating human sperm-egg recognition.","volume":"17","author":"Nixon","year":"2015","journal-title":"Asian J. Androl."},{"key":"B41","doi-asserted-by":"publisher","first-page":"160","DOI":"10.1093\/humupd\/6.2.160","article-title":"Sperm function assays and their predictive value for fertilization outcome in IVF therapy: a meta-analysis.","volume":"6","author":"Oehninger","year":"2000","journal-title":"Hum. Reprod. Update"},{"key":"B42","doi-asserted-by":"publisher","first-page":"1761","DOI":"10.1126\/science.1207438","article-title":"Human sperm binding is mediated by the sialyl-Lewis(x) oligosaccharide on the zona pellucida.","volume":"333","author":"Pang","year":"2011","journal-title":"Science"},{"key":"B43","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1093\/oxfordjournals.humrep.a136392","article-title":"Timing of in-vitro fertilization of cumulus-free and cumulus-enclosed human oocytes.","volume":"1","author":"Plachot","year":"1986","journal-title":"Hum. Reprod."},{"key":"B44","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1093\/molehr\/gas064","article-title":"Investigation of the mechanisms by which the molecular chaperone HSPA2 regulates the expression of sperm surface receptors involved in human sperm-oocyte recognition.","volume":"19","author":"Redgrove","year":"2013","journal-title":"Mol. Hum. Reprod."},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0050851","article-title":"The molecular chaperone HSPA2 plays a key role in regulating the expression of sperm surface receptors that mediate sperm-egg recognition.","volume":"7","author":"Redgrove","year":"2012","journal-title":"PLoS One"},{"key":"B46","doi-asserted-by":"publisher","first-page":"588","DOI":"10.2164\/jandrol.106.002279","article-title":"GM1 dynamics as a marker for membrane changes associated with the process of capacitation in murine and bovine spermatozoa.","volume":"28","author":"Selvaraj","year":"2007","journal-title":"J. Androl."},{"key":"B47","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1023\/A:1006951407168","article-title":"Cell surface galactosyltransferase: current issues.","volume":"15","author":"Shur","year":"1998","journal-title":"Glycoconj. J."},{"key":"B48","doi-asserted-by":"publisher","first-page":"137","DOI":"10.1016\/j.mce.2005.12.037","article-title":"Identification of novel gamete receptors that mediate sperm adhesion to the egg coat.","volume":"250","author":"Shur","year":"2006","journal-title":"Mol. Cell Endocrinol."},{"key":"B49","doi-asserted-by":"publisher","first-page":"843","DOI":"10.1095\/biolreprod42.5.843","article-title":"Human sperm plasma menbranes possess alpha-D-mannosidase activity but no galactosyltransferase activity but no falactosyltransferase activity.","volume":"42","author":"Tulsiani","year":"1990","journal-title":"Biol. Reprod."},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.3390\/cells10010133","article-title":"Ligands and receptors involved in the sperm-zona pellucida interactions in mammals.","volume":"10","author":"Tumova","year":"2021","journal-title":"Cells"},{"key":"B51","doi-asserted-by":"publisher","first-page":"583","DOI":"10.1093\/molehr\/gah200","article-title":"Capacitation-dependent concentration of lipid rafts in the apical ridge head area of porcine sperm cells.","volume":"11","author":"van Gestel","year":"2005","journal-title":"Mol. Hum. Reprod."},{"key":"B52","doi-asserted-by":"publisher","first-page":"1033","DOI":"10.1093\/biolre\/ioaa018","article-title":"LYPD4, mouse homolog of a human acrosome protein, is essential for sperm fertilizing ability and male fertility\u2020.","volume":"102","author":"Wang","year":"2020","journal-title":"Biol. Reprod."},{"key":"B53","doi-asserted-by":"publisher","first-page":"5750","DOI":"10.1021\/pr400753f","article-title":"Mapping of the N-linked glycoproteome of human spermatozoa.","volume":"12","author":"Wang","year":"2013","journal-title":"J. Proteome Res."},{"key":"B54","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1016\/S0092-8674(00)80558-9","article-title":"Mammalian fertilization: molecular aspects of gamete adhesion, exocytosis, and fusion.","volume":"96","author":"Wassarman","year":"1999","journal-title":"Cell"},{"key":"B55","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1038\/35055178","article-title":"A profile of fertilization in mammals.","volume":"3","author":"Wassarman","year":"2001","journal-title":"Nat. Cell. Biol."},{"key":"B56","first-page":"7","year":"2010","journal-title":"Laboratory Manual for the Examination and Processing of Human Semen."},{"key":"B57","doi-asserted-by":"publisher","first-page":"2541","DOI":"10.1093\/humrep\/13.9.2541","article-title":"Glycoproteins present in human follicular fluid that inhibit the zona-binding capacity of spermatozoa.","volume":"13","author":"Yao","year":"1998","journal-title":"Hum. Reprod."},{"key":"B58","doi-asserted-by":"publisher","first-page":"405","DOI":"10.1208\/s12248-014-9706-4","article-title":"Direct site-specific glycoform identification and quantitative comparison of glycoprotein therapeutics: imiglucerase and velaglucerase alfa.","volume":"17","author":"Ye","year":"2015","journal-title":"AAPS J."}],"container-title":["Frontiers in Cell and Developmental Biology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcell.2021.700396\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,7,20]],"date-time":"2021-07-20T10:15:53Z","timestamp":1626776153000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcell.2021.700396\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,20]]},"references-count":58,"alternative-id":["10.3389\/fcell.2021.700396"],"URL":"https:\/\/doi.org\/10.3389\/fcell.2021.700396","relation":{},"ISSN":["2296-634X"],"issn-type":[{"value":"2296-634X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,7,20]]},"article-number":"700396"}}