{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/25339"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/25339","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Fluorenscence depolarization study of internal tryptophan mobility in hydrated azurin films","abstract":"The effect of hydration on protein dynamics was investigated using steady state fluorescence depolarization on azurin, which has a single tryptophan residue well-buried in the hydrophobic interior. Azurin was imbedded in a thin, solid, water-permeable polymer film. This procedure inhibited whole-protein motion while allowing examination of internal degrees of rotational freedom in the protein as a function of film hydration. In the dry film at room temperature the tryptophan fluorescence emission showed the limiting polarization characteristic of low temperature solutions in which all motion is frozen. As the hydration was increased, the observed polarization value changed sharply at about 0.6 h (h =g water/g film), indicating an increase in the rotational mobility of the tryptophan. The fully hydrated film had a polarization equal to that of an azurin solution measurement extrapolated to infinite viscosity where the rotation of the protein as a whole 1s hindered. The conclusion is that the protein internal mobility is linked to the hydration level, and that hydration can turn on the full protein flexibility on the nanosecond time scale accessible by fluorescence techniques.","abstract_html":"The effect of hydration on protein dynamics was investigated using steady state fluorescence depolarization on azurin, which has a single tryptophan residue well-buried in the hydrophobic interior. Azurin was imbedded in a thin, solid, water-permeable polymer film. This procedure inhibited whole-protein motion while allowing examination of internal degrees of rotational freedom in the protein as a function of film hydration. In the dry film at room temperature the tryptophan fluorescence emission showed the limiting polarization characteristic of low temperature solutions in which all motion is frozen. As the hydration was increased, the observed polarization value changed sharply at about 0.6 h (h =g water/g film), indicating an increase in the rotational mobility of the tryptophan. The fully hydrated film had a polarization equal to that of an azurin solution measurement extrapolated to infinite viscosity where the rotation of the protein as a whole 1s hindered. The conclusion is that the protein internal mobility is linked to the hydration level, and that hydration can turn on the full protein flexibility on the nanosecond time scale accessible by fluorescence techniques.","abstract_has_math":false,"creators":["Limkeman, Mark Kenneth"],"institution":null,"degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Physics","degree_department":null,"school":null,"contributors":["Gratton, E."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2011,"date_issued":"2011-06-07T16:57:33Z","date_published":"2011-06-07T16:57:33Z","updated_at":"2026-07-22T22:25:24Z","subjects":["fluorescence depolarization","internal tryptophan mobility","hydrated azurin films","protein dynamics"],"languages":["en"],"rights":["1984 Mark Kenneth Limkeman"],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["826568"],"render_values":[{"text":"826568","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/25339","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Gratton, E."]},{"key":"dc:creator","label":"Author","values":["Limkeman, Mark Kenneth"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2011-06-07T16:57:33Z","10000-01-01","1984"]},{"key":"dc:type","label":"Dc Type","values":["Dissertation / Thesis","text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Physics"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["fluorescence depolarization","internal tryptophan mobility","hydrated azurin films","protein dynamics"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["1984 Mark Kenneth Limkeman"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["826568","http://hdl.handle.net/2142/25339"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["The effect of hydration on protein dynamics was investigated using steady state fluorescence depolarization on azurin, which has a single tryptophan residue well-buried in the hydrophobic interior. Azurin was imbedded in a thin, solid, water-permeable polymer film. This procedure inhibited whole-protein motion while allowing examination of internal degrees of rotational freedom in the protein as a function of film hydration. In the dry film at room temperature the tryptophan fluorescence emission showed the limiting polarization characteristic of low temperature solutions in which all motion is frozen. As the hydration was increased, the observed polarization value changed sharply at about 0.6 h (h =g water/g film), indicating an increase in the rotational mobility of the tryptophan. The fully hydrated film had a polarization equal to that of an azurin solution measurement extrapolated to infinite viscosity where the rotation of the protein as a whole 1s hindered. The conclusion is that the protein internal mobility is linked to the hydration level, and that hydration can turn on the full protein flexibility on the nanosecond time scale accessible by fluorescence techniques.","Submitted by Carolyn Mead (cmead2@illinois.edu) on 2011-06-07T16:57:33Z No. of bitstreams: 1 1984_limkeman.pdf: 3098975 bytes, checksum: 96e01c9f10526aa2225b42a1b4ff846d (MD5)","Made available in DSpace on 2011-06-07T16:57:33Z (GMT). No. of bitstreams: 1 1984_limkeman.pdf: 3098975 bytes, checksum: 96e01c9f10526aa2225b42a1b4ff846d (MD5) Previous issue date: 1984","Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Carolyn Mead (cmead2@illinois.edu) on 2011-06-07T16:57:33Z Item is restricted indefinitely.","Restriction data tranferred 2014-07-01T11:15:19-05:00 Original Data Group with Access UIUC Users [automated] Release Date: none Reason: Thesis","Thesis","U of I Only"]},{"key":"dc:title","label":"Title","values":["Fluorenscence depolarization study of internal tryptophan mobility in hydrated azurin films"]}]}],"canonical_facts":{"dc:contributor":["Gratton, E."],"dc:creator":["Limkeman, Mark Kenneth"],"dc:date":["2011-06-07T16:57:33Z","10000-01-01","1984"],"dc:description":["The effect of hydration on protein dynamics was investigated using steady state fluorescence depolarization on azurin, which has a single tryptophan residue well-buried in the hydrophobic interior. Azurin was imbedded in a thin, solid, water-permeable polymer film. This procedure inhibited whole-protein motion while allowing examination of internal degrees of rotational freedom in the protein as a function of film hydration. In the dry film at room temperature the tryptophan fluorescence emission showed the limiting polarization characteristic of low temperature solutions in which all motion is frozen. As the hydration was increased, the observed polarization value changed sharply at about 0.6 h (h =g water/g film), indicating an increase in the rotational mobility of the tryptophan. The fully hydrated film had a polarization equal to that of an azurin solution measurement extrapolated to infinite viscosity where the rotation of the protein as a whole 1s hindered. The conclusion is that the protein internal mobility is linked to the hydration level, and that hydration can turn on the full protein flexibility on the nanosecond time scale accessible by fluorescence techniques.","Submitted by Carolyn Mead (cmead2@illinois.edu) on 2011-06-07T16:57:33Z No. of bitstreams: 1 1984_limkeman.pdf: 3098975 bytes, checksum: 96e01c9f10526aa2225b42a1b4ff846d (MD5)","Made available in DSpace on 2011-06-07T16:57:33Z (GMT). No. of bitstreams: 1 1984_limkeman.pdf: 3098975 bytes, checksum: 96e01c9f10526aa2225b42a1b4ff846d (MD5) Previous issue date: 1984","Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Carolyn Mead (cmead2@illinois.edu) on 2011-06-07T16:57:33Z Item is restricted indefinitely.","Restriction data tranferred 2014-07-01T11:15:19-05:00 Original Data Group with Access UIUC Users [automated] Release Date: none Reason: Thesis","Thesis","U of I Only"],"dc:identifier":["826568","http://hdl.handle.net/2142/25339"],"dc:language":["en"],"dc:rights":["1984 Mark Kenneth Limkeman"],"dc:subject":["fluorescence depolarization","internal tryptophan mobility","hydrated azurin films","protein dynamics"],"dc:title":["Fluorenscence depolarization study of internal tryptophan mobility in hydrated azurin films"],"dc:type":["Dissertation / Thesis","text"],"thesis:degree_discipline":["Physics"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."]},"updated_at":"2026-07-22T22:25:24Z"}