{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/19904"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/19904","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"The role of BIP and PDI in the secretion of foreign proteins in the yeast Saccharomyces cerevisiae","abstract":"As a single-celled microbial eucaryotic host for protein expression, the yeast Saccharomyces cerevisiae offers some of the advantages of bacterial systems, such as ease of fermentation, and of eucaryotic systems, such as post-translational modifications of secreted proteins. One disadvantage, however, is that secretion of foreign proteins is generally inefficient.","abstract_html":"As a single-celled microbial eucaryotic host for protein expression, the yeast Saccharomyces cerevisiae offers some of the advantages of bacterial systems, such as ease of fermentation, and of eucaryotic systems, such as post-translational modifications of secreted proteins. One disadvantage, however, is that secretion of foreign proteins is generally inefficient.","abstract_has_math":false,"creators":["Robinson, Anne Skaja"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Chemical and Biomolecular Engineering","degree_department":null,"school":null,"contributors":["Lauffenburger, Douglas A."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2011,"date_issued":"2011-05-07T12:22:25Z","date_published":"2011-05-07T12:22:25Z","updated_at":"2026-07-22T22:25:14Z","subjects":["Biology, Cell","Chemistry, Biochemistry","Engineering, Chemical"],"languages":["eng"],"rights":["Copyright 1994 Robinson, Anne Skaja"],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["AAI9512530","(UMI)AAI9512530"],"render_values":[{"text":"AAI9512530","href":null,"code":true},{"text":"(UMI)AAI9512530","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/19904","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Lauffenburger, Douglas A."]},{"key":"dc:creator","label":"Author","values":["Robinson, Anne Skaja"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2011-05-07T12:22:25Z","10000-01-01","1994"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Chemical and Biomolecular Engineering"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Biology, Cell","Chemistry, Biochemistry","Engineering, Chemical"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 1994 Robinson, Anne Skaja"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["AAI9512530","(UMI)AAI9512530","http://hdl.handle.net/2142/19904"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["As a single-celled microbial eucaryotic host for protein expression, the yeast Saccharomyces cerevisiae offers some of the advantages of bacterial systems, such as ease of fermentation, and of eucaryotic systems, such as post-translational modifications of secreted proteins. One disadvantage, however, is that secretion of foreign proteins is generally inefficient.","The limiting step in protein secretion is often protein folding in the lumen of the endoplasmic reticulum (ER), a process assisted by accessory factors resident in this compartment. Chaperones, such as the hsp70 homolog binding protein (BiP), bind reversibly to the unfolded conformation of proteins, preventing irreversible aggregation. Foldases, such as protein disulfide isomerase (PDI), catalyze the formation and rearrangement of bonds which stabilize the folded conformation of proteins. I have examined the role of BiP and PDI in determining the efficiency of foreign protein secretion in the yeast Saccharomyces cerevisiae.","I have found that high-level expression of foreign genes does not always lead to increased production of foreign proteins. In fact, prolonged constitutive expression of foreign secreted proteins reduces soluble BiP and PDI protein to levels undetectable by Western immunoassay. Fifteen-fold overexpression of PDI from a strong glycolytic promoter results in significant enhancement of secretion for some foreign proteins. Improved secretion is correlated with decreased ER retention, indicative of accelerated folding. When the chromosomal copy of BiP is deleted, and BiP levels are tightly regulated from a plasmid-borne copy of the gene controlled by the CUP1 promoter, both secretion and growth are decreased significantly when BiP falls below wild type levels. A mechanistic model can account for the behavior of BiP experimentally, and predictions have been made for altering cellular properties to increase protein secretion.","Made available in DSpace on 2011-05-07T12:22:25Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9512530.pdf: 5154040 bytes, checksum: 53abbfdf1d3f08e9bf905a88959663a6 (MD5) Previous issue date: 1994","Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:40:13Z Item is restricted indefinitely.","Restriction data tranferred 2014-07-01T11:17:10-05:00 Original Data Group with Access UIUC Users [automated] Release Date: none Reason: ETDs are only available to UIUC Users without author permission","ETDs are only available to UIUC Users without author permission","U of I Only"]},{"key":"dc:title","label":"Title","values":["The role of BIP and PDI in the secretion of foreign proteins in the yeast Saccharomyces cerevisiae"]}]}],"canonical_facts":{"dc:contributor":["Lauffenburger, Douglas A."],"dc:creator":["Robinson, Anne Skaja"],"dc:date":["2011-05-07T12:22:25Z","10000-01-01","1994"],"dc:description":["As a single-celled microbial eucaryotic host for protein expression, the yeast Saccharomyces cerevisiae offers some of the advantages of bacterial systems, such as ease of fermentation, and of eucaryotic systems, such as post-translational modifications of secreted proteins. One disadvantage, however, is that secretion of foreign proteins is generally inefficient.","The limiting step in protein secretion is often protein folding in the lumen of the endoplasmic reticulum (ER), a process assisted by accessory factors resident in this compartment. Chaperones, such as the hsp70 homolog binding protein (BiP), bind reversibly to the unfolded conformation of proteins, preventing irreversible aggregation. Foldases, such as protein disulfide isomerase (PDI), catalyze the formation and rearrangement of bonds which stabilize the folded conformation of proteins. I have examined the role of BiP and PDI in determining the efficiency of foreign protein secretion in the yeast Saccharomyces cerevisiae.","I have found that high-level expression of foreign genes does not always lead to increased production of foreign proteins. In fact, prolonged constitutive expression of foreign secreted proteins reduces soluble BiP and PDI protein to levels undetectable by Western immunoassay. Fifteen-fold overexpression of PDI from a strong glycolytic promoter results in significant enhancement of secretion for some foreign proteins. Improved secretion is correlated with decreased ER retention, indicative of accelerated folding. When the chromosomal copy of BiP is deleted, and BiP levels are tightly regulated from a plasmid-borne copy of the gene controlled by the CUP1 promoter, both secretion and growth are decreased significantly when BiP falls below wild type levels. A mechanistic model can account for the behavior of BiP experimentally, and predictions have been made for altering cellular properties to increase protein secretion.","Made available in DSpace on 2011-05-07T12:22:25Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9512530.pdf: 5154040 bytes, checksum: 53abbfdf1d3f08e9bf905a88959663a6 (MD5) Previous issue date: 1994","Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:40:13Z Item is restricted indefinitely.","Restriction data tranferred 2014-07-01T11:17:10-05:00 Original Data Group with Access UIUC Users [automated] Release Date: none Reason: ETDs are only available to UIUC Users without author permission","ETDs are only available to UIUC Users without author permission","U of I Only"],"dc:identifier":["AAI9512530","(UMI)AAI9512530","http://hdl.handle.net/2142/19904"],"dc:language":["eng"],"dc:rights":["Copyright 1994 Robinson, Anne Skaja"],"dc:subject":["Biology, Cell","Chemistry, Biochemistry","Engineering, Chemical"],"dc:title":["The role of BIP and PDI in the secretion of foreign proteins in the yeast Saccharomyces cerevisiae"],"dc:type":["text"],"thesis:degree_discipline":["Chemical and Biomolecular Engineering"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:25:14Z"}