{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,12]],"date-time":"2026-03-12T07:35:19Z","timestamp":1773300919929,"version":"3.50.1"},"reference-count":56,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2014,8,7]],"date-time":"2014-08-07T00:00:00Z","timestamp":1407369600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000005","name":"Department of Defense","doi-asserted-by":"crossref","award":["W81XWH-11-1-0790"],"award-info":[{"award-number":["W81XWH-11-1-0790"]}],"id":[{"id":"10.13039\/100000005","id-type":"DOI","asserted-by":"crossref"}]},{"name":"National Science Foundation","award":["CHE: 1352122"],"award-info":[{"award-number":["CHE: 1352122"]}]},{"DOI":"10.13039\/100000928","name":"Welch Foundation","doi-asserted-by":"crossref","award":["AA-1854"],"award-info":[{"award-number":["AA-1854"]}],"id":[{"id":"10.13039\/100000928","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2014,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>This article utilized \u201cprotein charge ladders\u201d\u2014chemical derivatives of proteins with similar structure, but systematically altered net charge\u2014to quantify how missense mutations that cause amyotrophic lateral sclerosis (ALS) affect the net negative charge (Z) of superoxide dismutase\u20101 (SOD1) as a function of subcellular pH and Zn<jats:sup>2+<\/jats:sup> stoichiometry. Capillary electrophoresis revealed that the net charge of ALS\u2010variant SOD1 can be different in sign and in magnitude\u2014by up to 7.4 units per dimer at lysosomal pH\u2014than values predicted from standard p<jats:italic>K<\/jats:italic><jats:sub>a<\/jats:sub> values of amino acids and formal oxidation states of metal ions. At pH 7.4, the G85R, D90A, and G93R substitutions diminished the net negative charge of dimeric SOD1 by up to +2.29 units more than predicted; E100K lowered net charge by less than predicted. The binding of a single Zn<jats:sup>2+<\/jats:sup> to mutant SOD1 lowered its net charge by an additional +2.33\u2009\u00b1\u20090.01 to +3.18\u2009\u00b1\u20090.02 units, however, each protein regulated net charge when binding a second, third, or fourth Zn<jats:sup>2+<\/jats:sup> (\u0394<jats:italic>Z<\/jats:italic>\u2009&lt;\u20090.44\u2009\u00b1\u20090.07 per additional Zn<jats:sup>2+<\/jats:sup>). Both metalated and apo\u2010SOD1 regulated net charge across subcellular pH, without inverting from negative to positive at the theoretical pI. Differential scanning calorimetry, hydrogen\u2010deuterium exchange, and inductively coupled plasma mass spectrometry confirmed that the structure, stability, and metal content of mutant proteins were not significantly affected by lysine acetylation. Measured values of net charge should be used when correlating the biophysical properties of a specific ALS\u2010variant SOD1 protein with its observed aggregation propensity or clinical phenotype.<\/jats:p>","DOI":"10.1002\/pro.2526","type":"journal-article","created":{"date-parts":[[2014,7,23]],"date-time":"2014-07-23T03:42:56Z","timestamp":1406086976000},"page":"1417-1433","source":"Crossref","is-referenced-by-count":17,"title":["Protein charge ladders reveal that the net charge of ALS\u2010linked superoxide dismutase can be different in sign and magnitude from predicted values"],"prefix":"10.1002","volume":"23","author":[{"given":"Yunhua","family":"Shi","sequence":"first","affiliation":[{"name":"Department of Chemistry and Biochemistry Baylor University Waco Texas 76798\u20107348"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alireza","family":"Abdolvahabi","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry Baylor University Waco Texas 76798\u20107348"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bryan F.","family":"Shaw","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry Baylor University Waco Texas 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