{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,31]],"date-time":"2025-10-31T07:02:00Z","timestamp":1761894120416,"version":"build-2065373602"},"reference-count":33,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2005,4,13]],"date-time":"2005-04-13T00:00:00Z","timestamp":1113350400000},"content-version":"vor","delay-in-days":4485,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Electrophoresis"],"published-print":{"date-parts":[[1993,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Standard protocols for sample preparation for sodium dodecyl sulfate\u2010polyacrylamide gel electrophoresis (SDS\u2010PAGE) typically involve the combined use of heat and a reductant to fully disrupt protein\u2010protein interactions and allow for constant ratios of SDS\u2010binding to individual polypeptides. However, <jats:sup>14<\/jats:sup>C\u2010labeled forms of the membrane\u2010bound, active\u2010site\u2010containing 27 kDa polypeptide of ammonia monooxygenase from <jats:italic>Nitrosomonas europaea<\/jats:italic> undergo an aggregation reaction when cells or membranes are heated in the presence of SDS\u2010PAGE sample buffer. The aggregate produced after heating at 100\u00b0C is a soluble complex which fails to enter the stacking gel in discontinuous SDS\u2010PAGE gels. The extent of the aggregation reaction is dependent on the temperature of sample preparation, and the reaction exhibits first\u2010order kinetics at 65\u00b0C and 100\u00b0C (rates constants = 0.07 and 0.35 min<jats:sup>\u22121<\/jats:sup>, respectively). The rate of the aggregation reaction is further dependent on the concentration of reductant used in the sample buffer. However, the concentration of SDS does not significantly affect the rate of aggregation. The aggregated form of the 27 kDA polypeptide can be isolated by gel\u2010permeation chromatography in the presence of SDS. The aggregated protein can also be returned to the monomeric state by incubation at high pH in the presence of SDS. The aggregation reaction also occurs with <jats:sup>14<\/jats:sup>C<jats:sub>2<\/jats:sub>H<jats:sub>2<\/jats:sub>\u2010labeled polypeptides in other species of autotrophic nitrifiers and a methanotrophic bacterium which expresses the particulate form of methane monooxygenase. We conclude that strongly hydrophobic amino acid sequences present in ammonia monooxygenase are responsible for the aggregation phenomenon.<\/jats:p>","DOI":"10.1002\/elps.1150140197","type":"journal-article","created":{"date-parts":[[2005,5,28]],"date-time":"2005-05-28T09:32:37Z","timestamp":1117272757000},"page":"619-627","source":"Crossref","is-referenced-by-count":23,"title":["An electrophoretic study of the thermal\u2010 and reductant\u2010dependent aggregation of the 27 kDa component of ammonia monooxygenase from <i>Nitrosomonas europaea<\/i>"],"prefix":"10.1002","volume":"14","author":[{"given":"Michael R.","family":"Hyman","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel J.","family":"Arp","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2005,4,13]]},"reference":[{"key":"e_1_2_1_2_2","first-page":"39","volume-title":"Nitrification","author":"Wood P. 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