{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,6]],"date-time":"2025-11-06T20:04:54Z","timestamp":1762459494172,"version":"3.37.3"},"reference-count":15,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2017,6,6]],"date-time":"2017-06-06T00:00:00Z","timestamp":1496707200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"Juselius"},{"name":"Erkko"},{"name":"Finnish Academy foundations"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Neuropathology Appl Neurobio"],"published-print":{"date-parts":[[2018,8]]},"abstract":"Aims<\/jats:title>Previously, detection of ANO<\/jats:styled-content>5 protein has been complicated by unspecific antibodies, most of which have not identified the correct protein. The aims of the study were to specify ANO<\/jats:styled-content>5 protein expression in human skeletal muscle, and to investigate if the ANO<\/jats:styled-content>5 protein levels are affected by different ANO<\/jats:styled-content>5 mutations in anoctaminopathy patients.<\/jats:p><\/jats:sec>Methods<\/jats:title>Four different antibodies were tested for ANO<\/jats:styled-content>5 specificity. A sample preparation method compatible with membrane proteins, combined with tissue fractionation was used to determine ANO<\/jats:styled-content>5 expression in cell cultures expressing ANO<\/jats:styled-content>5, in normal muscles and eight patient biopsies with six different ANO<\/jats:styled-content>5 mutations in homozygous or compound heterozygous states, and in other dystrophies.<\/jats:p><\/jats:sec>Results<\/jats:title>Only one specific monoclonal N\u2010terminal ANO<\/jats:styled-content>5 antibody was efficient in detecting the protein, showing that ANO<\/jats:styled-content>5 is expressed as a single 107 kD<\/jats:styled-content> polypeptide in human skeletal muscle. The truncating mutations c.191dupA and c.1261C>T were found to abolish ANO<\/jats:styled-content>5 expression, whereas the studied point mutations had variable effects; however, all the ANO<\/jats:styled-content>5 mutations resulted in clearly reduced ANO<\/jats:styled-content>5 expression in the patient muscle membrane fraction. Attempts to detect ANO<\/jats:styled-content>5 using immunohistochemistry were not yet successful.<\/jats:bold><\/jats:p><\/jats:sec>Conclusions<\/jats:title>The data presented here indicate that the ANO<\/jats:styled-content>5 protein expression is decreased in ANO<\/jats:styled-content>5\u2010mutated muscular dystrophy and that most of the non\u2010truncating pathogenic ANO<\/jats:styled-content>5 mutations likely destabilize the protein and cause its degradation. The method described here allows direct analysis of human ANO<\/jats:styled-content>5 protein, which can be used in diagnostics, for evaluating the pathogenicity of the potentially harmful ANO<\/jats:styled-content>5 variants of uncertain significance.<\/jats:p><\/jats:sec>","DOI":"10.1111\/nan.12410","type":"journal-article","created":{"date-parts":[[2017,5,10]],"date-time":"2017-05-10T14:05:10Z","timestamp":1494425110000},"page":"441-448","source":"Crossref","is-referenced-by-count":18,"title":["Diagnostic anoctamin\u20105 protein defect in patients with ANO5\u2010mutated muscular dystrophy"],"prefix":"10.1111","volume":"44","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9249-8480","authenticated-orcid":false,"given":"A.","family":"Vihola","sequence":"first","affiliation":[{"name":"Folkh\u00e4lsan Institute of Genetics and Department of Medical Genetics Medicum University of Helsinki Helsinki 00014 Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"H.","family":"Luque","sequence":"additional","affiliation":[{"name":"Folkh\u00e4lsan Institute of Genetics and Department of Medical Genetics Medicum University of Helsinki Helsinki 00014 Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M.","family":"Savarese","sequence":"additional","affiliation":[{"name":"Folkh\u00e4lsan Institute of Genetics and Department of Medical Genetics Medicum University of Helsinki Helsinki 00014 Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S.","family":"Penttil\u00e4","sequence":"additional","affiliation":[{"name":"Neuromuscular Research Center University and University Hospital of Tampere Tampere Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M.","family":"Lindfors","sequence":"additional","affiliation":[{"name":"Neuromuscular Research Center University and University Hospital of Tampere Tampere Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"F.","family":"Leturcq","sequence":"additional","affiliation":[{"name":"Laboratoire de g\u00e9n\u00e9tique et biologie mol\u00e9culaire h\u00f4pital Cochin AP\u2010HP Universit\u00e9 Paris Descartes\u2010Sorbonne Paris Cit\u00e9 Paris France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"B.","family":"Eymard","sequence":"additional","affiliation":[{"name":"Institute of Myology Piti\u00e9\u2010Salp\u00eatri\u00e8re Hospital Paris France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0849-9144","authenticated-orcid":false,"given":"G.","family":"Tasca","sequence":"additional","affiliation":[{"name":"Istituto di Neurologia Universit\u00e0 Cattolica del Sacro Cuore Fondazione Policlinico Universitario \u201cA. 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