{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,12]],"date-time":"2026-03-12T05:08:31Z","timestamp":1773292111519,"version":"3.50.1"},"reference-count":27,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2003,2,24]],"date-time":"2003-02-24T00:00:00Z","timestamp":1046044800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["European Journal of Biochemistry"],"published-print":{"date-parts":[[2003,3]]},"abstract":"<jats:p>Human acid sphingomyelinase (haSMase, EC\u20033.1.4.12) catalyzes the lysosomal degradation of sphingomyelin to ceramide and phosphorylcholine. An inherited haSMase deficiency leads to Niemann\u2013Pick disease, a severe sphingolipid storage disorder.<\/jats:p><jats:p>The enzyme was purified and cloned over 10 years ago. Since then, only a few structural properties of haSMase have been elucidated. For understanding of its complex functions including its role in certain signaling and apoptosis events, complete structural information about the enzyme is necessary. Here, the identification of the disulfide bond pattern of haSMase is reported for the first time. Functional recombinant enzyme expressed in SF21 cells using the baculovirus expression system was purified and digested by trypsin. MALDI\u2010MS analysis of the resulting peptides revealed the four disulfide bonds Cys120\u2010Cys131, Cys385\u2010Cys431, Cys584\u2010Cys588 and Cys594\u2010Cys607. Two additional disulfide bonds (Cys221\u2010Cys226 and Cys227\u2010Cys250) which were not directly accessible by tryptic cleavage, were identified by a combination of a method of partial reduction and MALDI\u2010PSD analysis. In the sphingolipid activator protein (SAP)\u2010homologous N\u2010terminal domain of haSMase, one disulfide bond was assigned as Cys120\u2010Cys131. The existence of two additional disulfide bridges in this region was proved, as was expected for the known disulfide bond pattern of SAP\u2010type domains. These results support the hypothesis that haSMase possesses an intramolecular SAP\u2010type activator domain as predicted by sequence comparison [Ponting, C.P. (1994) <jats:italic>Protein Sci.<\/jats:italic>, <jats:bold>3<\/jats:bold>, 359\u2013361].<\/jats:p><jats:p>An additional analysis of haSMase isolated from human placenta shows that the recombinant and the native human protein possess an identical disulfide structure.<\/jats:p>","DOI":"10.1046\/j.1432-1033.2003.03435.x","type":"journal-article","created":{"date-parts":[[2004,12,23]],"date-time":"2004-12-23T02:53:30Z","timestamp":1103770410000},"page":"1076-1088","source":"Crossref","is-referenced-by-count":35,"title":["Human acid sphingomyelinase"],"prefix":"10.1111","volume":"270","author":[{"given":"Stephanie","family":"Lansmann","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christina G.","family":"Schuette","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Oliver","family":"Bartelsen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joerg","family":"Hoernschemeyer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thomas","family":"Linke","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Judith","family":"Weisgerber","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Konrad","family":"Sandhoff","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2003,2,24]]},"reference":[{"key":"e_1_2_37_2_2","first-page":"2601","volume-title":"The Metabolic and Molecular Bases of Inherited Disease","author":"Schuchman E.H.","year":"1995"},{"key":"e_1_2_37_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/0005-2760(87)90055-5"},{"key":"e_1_2_37_4_2","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1989.tb08382.x"},{"key":"e_1_2_37_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)93007-3"},{"key":"e_1_2_37_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(17)37705-0"},{"key":"e_1_2_37_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(96)01331-2"},{"key":"e_1_2_37_8_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1997.511_1a.x"},{"key":"e_1_2_37_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0955-0674(96)80061-5"},{"key":"e_1_2_37_10_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.274.5294.1855"},{"key":"e_1_2_37_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1388-1981(00)00042-1"},{"key":"e_1_2_37_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)61513-9"},{"key":"e_1_2_37_13_2","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.1994.1612"},{"key":"e_1_2_37_14_2","doi-asserted-by":"publisher","DOI":"10.1002\/pro.5560030219"},{"key":"e_1_2_37_15_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.270.17.9953"},{"key":"e_1_2_37_16_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1432-1327.1999.00521.x"},{"key":"e_1_2_37_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-1656(98)00070-4"},{"key":"e_1_2_37_18_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2222.1998.00312.x"},{"key":"e_1_2_37_19_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac00093a030"},{"key":"e_1_2_37_20_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00414a041"},{"key":"e_1_2_37_21_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bi.54.070185.003215"},{"key":"e_1_2_37_22_2","doi-asserted-by":"publisher","DOI":"10.1002\/pro.5560060215"},{"key":"e_1_2_37_23_2","doi-asserted-by":"publisher","DOI":"10.1002\/pro.5560070421"},{"key":"e_1_2_37_24_2","first-page":"137","article-title":"Sequencing of peptides in a time\u2010of\u2010flight mass spectrometer: Evaluation of post source decay following matrix\u2010assisted laser desorption ionization (MALDI)","volume":"86","author":"Kaufmann R.","year":"1994","journal-title":"Int. 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