{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T15:05:37Z","timestamp":1787670337151,"version":"build-2736575974"},"reference-count":59,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2024,1,23]],"date-time":"2024-01-23T00:00:00Z","timestamp":1705968000000},"content-version":"am","delay-in-days":336,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#am"},{"start":{"date-parts":[[2023,2,21]],"date-time":"2023-02-21T00:00:00Z","timestamp":1676937600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","award":["MCB 1410854"],"award-info":[{"award-number":["MCB 1410854"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","award":["MCB 1909664"],"award-info":[{"award-number":["MCB 1909664"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2023,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Proteins in the cellular milieu reside in environments crowded by macromolecules and other solutes. Although crowding can significantly impact the protein folded state stability, most experiments are conducted in dilute buffered solutions. To resolve the effect of crowding on protein stability, we use\n                    <jats:sup>19<\/jats:sup>\n                    F nuclear magnetic resonance spectroscopy to follow the reversible, two\u2010state unfolding thermodynamics of the N\u2010terminal Src homology 3 domain of the\n                    <jats:italic>Drosophila<\/jats:italic>\n                    signal transduction protein drk in the presence of polyethylene glycols (PEGs) of various molecular weights and concentrations. Contrary to most current theories of crowding that emphasize steric protein\u2013crowder interactions as the main driving force for entropically favored stabilization, our experiments show that PEG stabilization is accompanied by significant heat release, and entropy disfavors folding. Using our newly developed model, we find that stabilization by ethylene glycol and small PEGs is driven by favorable binding to the folded state. In contrast, for larger PEGs, chemical or soft PEG\u2013protein interactions do not play a significant role. Instead, folding is favored by excluded volume PEG\u2013protein interactions and an exothermic nonideal mixing contribution from release of confined PEG and water upon folding. Our results indicate that crowding acts through molecular interactions subtler than previously assumed and that interactions between solution components with both the folded and unfolded states must be carefully considered.\n                  <\/jats:p>","DOI":"10.1002\/pro.4573","type":"journal-article","created":{"date-parts":[[2023,1,24]],"date-time":"2023-01-24T02:55:49Z","timestamp":1674528949000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":60,"title":["Resolving the enthalpy of protein stabilization by macromolecular crowding"],"prefix":"10.1002","volume":"32","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3071-3322","authenticated-orcid":false,"given":"Claire J.","family":"Stewart","sequence":"first","affiliation":[{"name":"Department of Chemistry University of North Carolina at Chapel Hill Chapel Hill North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4514-5516","authenticated-orcid":false,"given":"Gil I.","family":"Olgenblum","sequence":"additional","affiliation":[{"name":"Institute of Chemistry &amp; the Fritz Haber Research Center, The Hebrew University Jerusalem Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashlee","family":"Propst","sequence":"additional","affiliation":[{"name":"Department of Chemistry University of North Carolina at Chapel Hill Chapel Hill North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3057-9485","authenticated-orcid":false,"given":"Daniel","family":"Harries","sequence":"additional","affiliation":[{"name":"Institute of Chemistry &amp; the Fritz Haber Research Center, The Hebrew University Jerusalem 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