{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T13:08:29Z","timestamp":1788786509124,"version":"build-2803163510"},"reference-count":56,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T00:00:00Z","timestamp":1771286400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T00:00:00Z","timestamp":1771286400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2026,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The cytoplasm of a cell is inherently crowded with diverse biological macromolecules of different sizes, shapes, and compositions. These macromolecular crowders strongly influence the thermodynamics and kinetics of essential biological processes, including protein folding, association, and enzymatic activity. Therefore, in our earlier study, we examined PEG size\u2010dependent crowding effects on protein stability via water\u2010network modulation; however, excluded\u2010volume (entropic) contributions varied and were not systematically controlled. To address this limitation, in this work, we examined the shape\u2010dependent effects of crowding by selecting similarly sized crowders, PEG\u201035, Ficoll\u201070, and dextran\u201040, to mimic the heterogeneous shapes of crystallins in the eye lens. Our results revealed distinct outcomes: PEG\u201035 facilitated the thermal unfolding of H\u03b3DC, dextran\u201040 counteracted unfolding, while Ficoll\u201070 produced no significant effect. The thermodynamic analysis provided even more intriguing insights. Both PEG\u201035 and Ficoll\u201070 entropically destabilized H\u03b3DC, in direct contrast to the predictions of traditional excluded volume theory, whereas dextran\u201040 exerted an entropic stabilizing effect. To rationalize these observations, we hypothesized the associated water stabilization mechanism (AWSM), which emphasizes the critical role of hydration water structure in protein stability under crowded conditions. Collectively, our findings highlight the importance of associated water in shaping protein stability and solubility, offering new perspectives on shape\u2010dependent crowding effects. The results not only account for entropic destabilization and entropy\u2013enthalpy compensation, but also demonstrate that the thick, resilient hydration shell surrounding human \u03b3\u2010D crystallin underpins its long\u2010term stability and solubility within the dense and crowded environment of the eye lens.<\/jats:p>","DOI":"10.1002\/pro.70507","type":"journal-article","created":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T12:18:59Z","timestamp":1771330739000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Role of associated water in stabilizing human \u03b3\u2010D crystallin under shape\u2010dependent macromolecular crowding"],"prefix":"10.1002","volume":"35","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-1384-441X","authenticated-orcid":false,"given":"Bhupendra","family":"Singh","sequence":"first","affiliation":[{"name":"Department of Chemistry Indian Institute of Technology Kanpur  Kanpur UP India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8202-1854","authenticated-orcid":false,"given":"Pratik","family":"Sen","sequence":"additional","affiliation":[{"name":"Department of Chemistry Indian Institute of Technology Kanpur  Kanpur UP India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,2,17]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2010.05.067"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1021\/cr020661"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1610541113"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2018.01.011"},{"key":"e_1_2_10_6_1","first-page":"415","article-title":"Mechanism of human \u03b3D\u2010crystallin protein aggregation in UV\u2010C light","volume":"27","author":"Bawankar M","year":"2021","journal-title":"Mol 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