{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T05:07:27Z","timestamp":1789967247914,"version":"4.0.1"},"reference-count":37,"publisher":"Portland Press Ltd.","issue":"6","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2017,3,15]]},"abstract":"<jats:p>Sperm-specific phospholipase C zeta (PLC\u03b6) is widely considered to be the physiological stimulus that evokes intracellular calcium (Ca2+) oscillations that are essential for the initiation of egg activation during mammalian fertilisation. A recent genetic study reported a male infertility case that was directly associated with a point mutation in the PLC\u03b6 C2 domain, where an isoleucine residue had been substituted with a phenylalanine (I489F). Here, we have analysed the effect of this mutation on the in vivo Ca2+ oscillation-inducing activity and the in vitro biochemical properties of human PLC\u03b6. Microinjection of cRNA or recombinant protein corresponding to PLC\u03b6I489F mutant at physiological concentrations completely failed to cause Ca2+ oscillations and trigger development. However, this infertile phenotype could be effectively rescued by microinjection of relatively high (non-physiological) amounts of recombinant mutant PLC\u03b6I489F protein, leading to Ca2+ oscillations and egg activation. Our in vitro biochemical analysis suggested that the PLC\u03b6I489F mutant displayed similar enzymatic properties, but dramatically reduced binding to PI(3)P and PI(5)P-containing liposomes compared with wild-type PLC\u03b6. Our findings highlight the importance of PLC\u03b6 at fertilisation and the vital role of the C2 domain in PLC\u03b6 function, possibly due to its novel binding characteristics.<\/jats:p>","DOI":"10.1042\/bcj20161057","type":"journal-article","created":{"date-parts":[[2017,3,8]],"date-time":"2017-03-08T01:20:23Z","timestamp":1488936023000},"page":"1003-1016","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":35,"title":["Male infertility-linked point mutation reveals a vital binding role for the C2 domain of sperm PLC\u03b6"],"prefix":"10.1042","volume":"474","author":[{"given":"Michail","family":"Nomikos","sequence":"first","affiliation":[{"name":"College of Medicine, Qatar University, PO Box 2713, Doha, Qatar"},{"name":"College of Biomedical and Life Sciences, School of Biosciences, Cardiff University, Cardiff, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Panagiotis","family":"Stamatiadis","sequence":"additional","affiliation":[{"name":"College of Biomedical and Life Sciences, School of Biosciences, Cardiff University, Cardiff, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica R.","family":"Sanders","sequence":"additional","affiliation":[{"name":"College of Biomedical and Life Sciences, School of Biosciences, Cardiff University, Cardiff, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Konrad","family":"Beck","sequence":"additional","affiliation":[{"name":"College of Biomedical and Life Sciences, School of Dentistry, Cardiff University, Cardiff, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian L.","family":"Calver","sequence":"additional","affiliation":[{"name":"College of Biomedical and Life Sciences, School of Biosciences, Cardiff University, Cardiff, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luke","family":"Buntwal","sequence":"additional","affiliation":[{"name":"College of Biomedical and Life Sciences, School of Biosciences, Cardiff University, Cardiff, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Morgan","family":"Lofty","sequence":"additional","affiliation":[{"name":"College of Biomedical and Life Sciences, School of Biosciences, Cardiff University, Cardiff, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zili","family":"Sideratou","sequence":"additional","affiliation":[{"name":"National Center for Scientific Research \u2018Demokritos\u2019, 15310 Aghia Paraskevi, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karl","family":"Swann","sequence":"additional","affiliation":[{"name":"College of Biomedical and Life Sciences, School of Biosciences, Cardiff University, Cardiff, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"F. Anthony","family":"Lai","sequence":"additional","affiliation":[{"name":"College of Biomedical and Life Sciences, School of Biosciences, Cardiff University, Cardiff, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","published-online":{"date-parts":[[2017,3,7]]},"reference":[{"key":"2021112216461110900_BCJ-2016-1057C1","doi-asserted-by":"publisher","first-page":"157","DOI":"10.1006\/dbio.1999.9340","article-title":"Comparative biology of calcium signaling during fertilization and egg activation in animals","volume":"211","author":"Stricker","year":"1999","journal-title":"Dev. Biol."},{"key":"2021112216461110900_BCJ-2016-1057C2","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1006\/dbio.2002.0600","article-title":"Egg activation at fertilization: where it all begins","volume":"245","author":"Runft","year":"2002","journal-title":"Dev. Biol."},{"key":"2021112216461110900_BCJ-2016-1057C3","doi-asserted-by":"publisher","first-page":"3609","DOI":"10.1016\/j.febslet.2013.10.008","article-title":"Sperm PLC\u03b6: from structure to Ca2+ oscillations, egg activation and therapeutic potential","volume":"587","author":"Nomikos","year":"2013","journal-title":"FEBS Lett."},{"key":"2021112216461110900_BCJ-2016-1057C4","doi-asserted-by":"crossref","first-page":"3533","DOI":"10.1242\/dev.129.15.3533","article-title":"PLC\u03b6: a sperm-specific trigger of Ca(2+) oscillations in eggs and embryo development","volume":"129","author":"Saunders","year":"2002","journal-title":"Development"},{"key":"2021112216461110900_BCJ-2016-1057C5","doi-asserted-by":"publisher","first-page":"76","DOI":"10.1016\/j.fertnstert.2012.08.035","article-title":"Phospholipase C\u03b6 rescues failed oocyte activation in a prototype of male factor infertility","volume":"99","author":"Nomikos","year":"2013","journal-title":"Fertil. Steril."},{"key":"2021112216461110900_BCJ-2016-1057C6","doi-asserted-by":"publisher","first-page":"702","DOI":"10.1093\/molehr\/gav034","article-title":"Functional disparity between human PAWP and PLC\u03b6 in the generation of Ca2+ oscillations for oocyte activation","volume":"21","author":"Nomikos","year":"2015","journal-title":"Mol. Hum. Reprod."},{"key":"2021112216461110900_BCJ-2016-1057C7","doi-asserted-by":"publisher","first-page":"94","DOI":"10.1095\/biolreprod.115.131441","article-title":"Sperm postacrosomal WW domain-binding protein is not required for mouse egg activation","volume":"93","author":"Satouh","year":"2015","journal-title":"Biol. Reprod."},{"key":"2021112216461110900_BCJ-2016-1057C8","doi-asserted-by":"publisher","first-page":"878","DOI":"10.1093\/hmg\/ddv617","article-title":"Homozygous mutation of PLCZ1 leads to defective human oocyte activation and infertility that is not rescued by the WW-binding protein PAWP","volume":"25","author":"Escoffier","year":"2016","journal-title":"Hum. Mol. Genet."},{"key":"2021112216461110900_BCJ-2016-1057C9","doi-asserted-by":"publisher","first-page":"371","DOI":"10.1042\/BST20140291","article-title":"Novel signalling mechanism and clinical applications of sperm-specific PLC\u03b6","volume":"43","author":"Nomikos","year":"2015","journal-title":"Biochem. Soc. Trans."},{"key":"2021112216461110900_BCJ-2016-1057C10","doi-asserted-by":"publisher","first-page":"29519","DOI":"10.1074\/jbc.M115.658443","article-title":"Essential role of the EF-hand domain in targeting sperm phospholipase C\u03b6 to membrane phosphatidylinositol 4,5-bisphosphate (PIP2)","volume":"290","author":"Nomikos","year":"2015","journal-title":"J. Biol. Chem."},{"key":"2021112216461110900_BCJ-2016-1057C11","doi-asserted-by":"publisher","first-page":"3671","DOI":"10.1172\/JCI36942","article-title":"Human sperm devoid of PLC, zeta 1 fail to induce Ca(2+) release and are unable to initiate the first step of embryo development","volume":"118","author":"Yoon","year":"2008","journal-title":"J. Clin. Invest."},{"key":"2021112216461110900_BCJ-2016-1057C12","doi-asserted-by":"publisher","first-page":"2417","DOI":"10.1093\/humrep\/dep207","article-title":"Reduced amounts and abnormal forms of phospholipase C zeta (PLC\u03b6) in spermatozoa from infertile men","volume":"24","author":"Heytens","year":"2009","journal-title":"Hum. Reprod."},{"key":"2021112216461110900_BCJ-2016-1057C13","doi-asserted-by":"publisher","first-page":"690","DOI":"10.1093\/humupd\/dmq018","article-title":"Oocyte activation, phospholipase C zeta and human infertility","volume":"16","author":"Kashir","year":"2010","journal-title":"Hum. Reprod. Update"},{"key":"2021112216461110900_BCJ-2016-1057C14","doi-asserted-by":"publisher","first-page":"222","DOI":"10.1093\/humrep\/der384","article-title":"A maternally inherited autosomal point mutation in human phospholipase C zeta (PLC\u03b6) leads to male infertility","volume":"27","author":"Kashir","year":"2012","journal-title":"Hum. Reprod."},{"key":"2021112216461110900_BCJ-2016-1057C15","doi-asserted-by":"publisher","first-page":"211","DOI":"10.1042\/BJ20101772","article-title":"Male infertility-linked point mutation disrupts the Ca2+ oscillation-inducing and PIP2 hydrolysis activity of sperm PLC\u03b6","volume":"434","author":"Nomikos","year":"2011","journal-title":"Biochem. J."},{"key":"2021112216461110900_BCJ-2016-1057C16","doi-asserted-by":"publisher","first-page":"1273","DOI":"10.1016\/S0093-691X(00)00271-5","article-title":"Activation of bovine oocytes following intracytoplasmic sperm injection (ICSI)","volume":"53","author":"Chung","year":"2000","journal-title":"Theriogenology"},{"key":"2021112216461110900_BCJ-2016-1057C17","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1007\/978-1-59745-202-1_2","article-title":"Use of luciferase chimaera to monitor PLC\u03b6 expression in mouse eggs","volume":"518","author":"Swann","year":"2009","journal-title":"Methods Mol. Biol."},{"key":"2021112216461110900_BCJ-2016-1057C18","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1016\/j.ydbio.2006.06.032","article-title":"Ca2+ oscillations stimulate an ATP increase during fertilization of mouse eggs","volume":"298","author":"Campbell","year":"2006","journal-title":"Dev. Biol."},{"key":"2021112216461110900_BCJ-2016-1057C19","doi-asserted-by":"publisher","first-page":"2411","DOI":"10.1002\/pro.5560041120","article-title":"How to measure and predict the molar absorption coefficient of a protein","volume":"4","author":"Pace","year":"1995","journal-title":"Protein Sci."},{"key":"2021112216461110900_BCJ-2016-1057C20","doi-asserted-by":"publisher","first-page":"307","DOI":"10.1002\/(SICI)1097-0134(19990515)35:3&lt;307::AID-PROT4&gt;3.0.CO;2-3","article-title":"Analyzing protein circular dichroism spectra for accurate secondary structures","volume":"35","author":"Johnson","year":"1999","journal-title":"Proteins Struct. Funct. Genet."},{"key":"2021112216461110900_BCJ-2016-1057C21","doi-asserted-by":"publisher","first-page":"392","DOI":"10.1002\/bip.20853","article-title":"Protein secondary structure analyses from circular dichroism spectroscopy: methods and reference databases","volume":"89","author":"Whitmore","year":"2008","journal-title":"Biopolymers"},{"key":"2021112216461110900_BCJ-2016-1057C22","doi-asserted-by":"publisher","first-page":"1630","DOI":"10.1093\/bioinformatics\/btr234","article-title":"A reference dataset for the analyses of membrane protein secondary structures and transmembrane residues using circular dichroism spectroscopy","volume":"27","author":"Abdul-Gader","year":"2011","journal-title":"Bioinformatics"},{"key":"2021112216461110900_BCJ-2016-1057C23","doi-asserted-by":"publisher","first-page":"282","DOI":"10.1016\/S0076-6879(04)83012-X","article-title":"Analysis of circular dichroism data","volume":"383","author":"Greenfield","year":"2004","journal-title":"Methods Enzymol."},{"key":"2021112216461110900_BCJ-2016-1057C24","doi-asserted-by":"publisher","first-page":"W252","DOI":"10.1093\/nar\/gku340","article-title":"SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information","volume":"42","author":"Biasini","year":"2014","journal-title":"Nucleic Acids Res."},{"key":"2021112216461110900_BCJ-2016-1057C25","doi-asserted-by":"publisher","first-page":"427","DOI":"10.1042\/BJ20110953","article-title":"Novel regulation of PLC\u03b6 activity via its XY-linker","volume":"438","author":"Nomikos","year":"2011","journal-title":"Biochem. J."},{"key":"2021112216461110900_BCJ-2016-1057C26","doi-asserted-by":"publisher","first-page":"852","DOI":"10.1093\/molehr\/gat070","article-title":"Chimeras of sperm PLC\u03b6 reveal disparate protein domain functions in the generation of intracellular Ca2+ oscillations in mammalian eggs at fertilization","volume":"19","author":"Theodoridou","year":"2013","journal-title":"Mol. Hum. Reprod."},{"key":"2021112216461110900_BCJ-2016-1057C27","doi-asserted-by":"publisher","first-page":"31011","DOI":"10.1074\/jbc.M500629200","article-title":"Role of phospholipase C-\u03b6 domains in Ca2+-dependent phosphatidylinositol 4,5-bisphosphate hydrolysis and cytoplasmic Ca2+ oscillations","volume":"280","author":"Nomikos","year":"2005","journal-title":"J. Biol. Chem."},{"key":"2021112216461110900_BCJ-2016-1057C28","doi-asserted-by":"publisher","first-page":"585","DOI":"10.1387\/ijdb.072530ks","article-title":"The dynamics of calcium oscillations that activate mammalian eggs","volume":"52","author":"Swann","year":"2008","journal-title":"Int. J. Dev. Biol."},{"key":"2021112216461110900_BCJ-2016-1057C29","doi-asserted-by":"publisher","first-page":"3608","DOI":"10.1021\/bi0360509","article-title":"Effect of signal peptide on the stability and folding kinetics of maltose binding protein","volume":"43","author":"Beena","year":"2004","journal-title":"Biochemistry"},{"key":"2021112216461110900_BCJ-2016-1057C30","doi-asserted-by":"publisher","first-page":"489","DOI":"10.1093\/molehr\/gau011","article-title":"Human PLC\u03b6 exhibits superior fertilization potency over mouse PLC\u03b6 in triggering the Ca2+ oscillations required for mammalian oocyte activation","volume":"20","author":"Nomikos","year":"2014","journal-title":"Mol. Hum. Reprod."},{"key":"2021112216461110900_BCJ-2016-1057C31","doi-asserted-by":"publisher","first-page":"21015","DOI":"10.1074\/jbc.M412123200","article-title":"The role of EF-hand domains and C2 domain in regulation of enzymatic activity of phospholipase C\u03b6","volume":"280","author":"Kouchi","year":"2005","journal-title":"J. Biol. Chem."},{"key":"2021112216461110900_BCJ-2016-1057C32","doi-asserted-by":"publisher","first-page":"2582","DOI":"10.1242\/jcs.083485","article-title":"Phospholipase C\u03b6 binding to PtdIns(4,5)P2 requires the XY-linker region","volume":"124","author":"Nomikos","year":"2011","journal-title":"J. Cell Sci."},{"key":"2021112216461110900_BCJ-2016-1057C33","doi-asserted-by":"publisher","first-page":"611","DOI":"10.1530\/rep.0.1240611","article-title":"Sperm phospholipase C zeta from humans and cynomolgus monkeys triggers Ca2+ oscillations, activation and development of mouse oocytes","volume":"124","author":"Cox","year":"2002","journal-title":"Reproduction"},{"key":"2021112216461110900_BCJ-2016-1057C34","doi-asserted-by":"publisher","first-page":"992","DOI":"10.1095\/biolreprod.104.036244","article-title":"Transgenic RNA interference reveals role for mouse sperm phospholipase C\u03b6 in triggering Ca2+ oscillations during fertilization","volume":"72","author":"Knott","year":"2005","journal-title":"Biol. Reprod."},{"key":"2021112216461110900_BCJ-2016-1057C35","doi-asserted-by":"publisher","first-page":"1204","DOI":"10.1016\/j.bbrc.2014.04.078","article-title":"Sperm-induced Ca2+ release during egg activation in mammals","volume":"450","author":"Kashir","year":"2014","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"2021112216461110900_BCJ-2016-1057C36","doi-asserted-by":"publisher","first-page":"58","DOI":"10.1016\/j.jbior.2015.11.009","article-title":"Phospholipase C zeta (PLC\u03b6) and male infertility: clinical update and topical developments","volume":"61","author":"Amdani","year":"2016","journal-title":"Adv. Biol. Regul."},{"key":"2021112216461110900_BCJ-2016-1057C37","doi-asserted-by":"publisher","first-page":"10408","DOI":"10.1074\/jbc.M313801200","article-title":"Recombinant phospholipase C\u03b6 has high Ca2+ sensitivity and induces Ca2+ oscillations in mouse eggs","volume":"279","author":"Kouchi","year":"2004","journal-title":"J. Biol. Chem."}],"container-title":["Biochemical Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/474\/6\/1003\/689341\/bcj-2016-1057.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/474\/6\/1003\/689341\/bcj-2016-1057.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,22]],"date-time":"2021-11-22T23:30:03Z","timestamp":1637623803000},"score":1,"resource":{"primary":{"URL":"https:\/\/portlandpress.com\/biochemj\/article\/474\/6\/1003\/49603\/Male-infertility-linked-point-mutation-reveals-a"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,3,7]]},"references-count":37,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2017,3,15]]}},"URL":"https:\/\/doi.org\/10.1042\/bcj20161057","relation":{},"ISSN":["0264-6021","1470-8728"],"issn-type":[{"value":"0264-6021","type":"print"},{"value":"1470-8728","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,3,7]]}}}