{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,18]],"date-time":"2026-01-18T13:40:21Z","timestamp":1768743621137,"version":"3.49.0"},"reference-count":41,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2008,12,31]],"date-time":"2008-12-31T00:00:00Z","timestamp":1230681600000},"content-version":"vor","delay-in-days":6027,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[1992,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>An enzymatic procedure for the complete removal of the <jats:italic>N<\/jats:italic>\u2010linked and <jats:italic>O<\/jats:italic>\u2010linked oligosaccharide side chains of the sex steroid\u2010binding proteins (SBP or SHBG) of human and rabbit plasma under native conditions is described. Deglycosylation was catalyzed by <jats:italic>N<\/jats:italic>\u2010glycanase, neuraminidase, and <jats:italic>O<\/jats:italic>\u2010glycanase and was monitored by SDS\u2010PAGE, lectin blotting, and molecular weight analyses by electrospray mass spectrometry. Digestion of rabbit SBP with <jats:italic>N<\/jats:italic>\u2010glycanase generated a major 39,777\u2010Da protein and two minor ones of 39,389 and 39,545 Da. The molecular weight of the major protein agrees with the molecular weight calculated from the sequence of the sugar\u2010free polypeptide monomer (39,769 Da: Griffin, P.R., Kumar, S., Shabanowitz, J., Charbonneau, H., Namkung, P.C., Walsh, K.A., Hunt, D.F., &amp; Petra, P.H., 1989, <jats:italic>J. Biol. Chem. 264<\/jats:italic>, 19066\u201319075), whereas the other two are deglycosylated proteolytic cleavage products lacking the TQR and TQ sequences at the amino\u2010terminus. The N\u2010 and <jats:italic>O<\/jats:italic>\u2010linked side chains of human SBP were removed by sequential digestion with <jats:italic>N<\/jats:italic>\u2010glycanase and neuraminidase\/<jats:italic>O<\/jats:italic>\u2010glycanase. A 38,771\u2010Da protein was generated, which agrees well with the molecular weight of the sugar\u2010free polypeptide monomer (Walsh, K.A., Titani, K., Kumar, S., Hayes, R., &amp; Petra, P.H., 1986, <jats:italic>Biochemistry 25<\/jats:italic>, 7584\u20137590). <jats:italic>N<\/jats:italic>\u2010deglycosylation of human and rabbit SBP has no effect on the steroid\u2010binding activity, but removal of the <jats:italic>O<\/jats:italic>\u2010linked side chains of <jats:italic>N<\/jats:italic>\u2010deglycosylated human SBP results in an apparent 50% loss of steroid\u2010binding activity and an increase in the <jats:italic>K<jats:sub>d<\/jats:sub><\/jats:italic> for the binding of 5<jats:italic>\u03b1<\/jats:italic>\u2010dihydrotestosterone from 0.3 nM to 0.9 nM. There are no changes in steroid\u2010binding specificity. The apparent loss of activity of <jats:italic>O<\/jats:italic>\u2010deglycosylated human SBP is probably due to the small changes in the <jats:italic>K<jats:sub>d<\/jats:sub><\/jats:italic>, which could influence the equilibrium concentration of bound SBP when measured under standard assay conditions. We conclude that deglycosylation has very little effect on steroid\u2010binding activity and that the oligosaccharide side chains must serve other functions in the physiology of SBP.<\/jats:p>","DOI":"10.1002\/pro.5560010708","type":"journal-article","created":{"date-parts":[[2009,2,9]],"date-time":"2009-02-09T23:39:33Z","timestamp":1234222773000},"page":"902-909","source":"Crossref","is-referenced-by-count":23,"title":["Complete enzymatic deglycosylation of native sex steroid\u2010binding protein (SBP or SHBG) of human and rabbit plasma: Effect on the steroid\u2010binding activity"],"prefix":"10.1002","volume":"1","author":[{"given":"Philip H.","family":"Petra","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wei","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Patrick R.","family":"Griffin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"suffix":"III","given":"John R.","family":"Yates","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Katherine","family":"Moore","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2008,12,31]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/0304-4165(83)90130-7"},{"key":"e_1_2_1_3_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI105588"},{"key":"e_1_2_1_4_1","doi-asserted-by":"publisher","DOI":"10.1210\/mend-5-11-1723"},{"key":"e_1_2_1_5_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.77.10.5678"},{"key":"e_1_2_1_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022-4731(82)90641-0"},{"key":"e_1_2_1_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022-4731(85)90152-9"},{"key":"e_1_2_1_8_1","doi-asserted-by":"publisher","DOI":"10.1210\/endo-124-6-2809"},{"key":"e_1_2_1_9_1","doi-asserted-by":"crossref","first-page":"8430","DOI":"10.1016\/S0021-9258(18)47582-5","article-title":"Subunits of human sex hormone binding globulin. 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