Multicopy Overexpression of Bovine Pancreatic Trypsin Inhibitor Saturates the Protein Folding and Secretory Capacity of Saccharomyces cerevisiae
Identifieur interne : 001843 ( Istex/Checkpoint ); précédent : 001842; suivant : 001844Multicopy Overexpression of Bovine Pancreatic Trypsin Inhibitor Saturates the Protein Folding and Secretory Capacity of Saccharomyces cerevisiae
Auteurs : R. Parekh [États-Unis] ; K. Forrester [États-Unis] ; D. Wittrup [États-Unis]Source :
- Protein Expression and Purification [ 1046-5928 ] ; 1995.
Abstract
Abstract: Bovine pancreatic trypsin inhibitor (BPTI) was expressed and secreted from a synthetic gene as a model system for the study of protein folding and secretion in Saccharomyces cerevisiae. The efficiency of different leader sequences in directing BPTI secretion was examined, and up to 11 μg/ml of active BPTI was secreted. In some fusion constructs, inefficient proteolytic processing by Kex2p, Ste13p, and signal peptidase were observed immediately adjacent to the BPTI N terminus. Insertion of dipeptide spacers improved endoproteolytic processing substantially but the level of secretion was unchanged, Overexpression from a 2-μm multicopy vector results in essentially unchanged BPTI secretion as compared to expression from a single copy centromere vector. BPTI expressed from a multicopy vector accumulates intracellularly in an unfolded form, indicating that available secretory chaperones and foldases can be saturated by increasing the rate of BPTI synthesis.
Url:
DOI: 10.1006/prep.1995.1071
Affiliations:
Links toward previous steps (curation, corpus...)
Links to Exploration step
ISTEX:3EC9F7D7DAC6F7402AEDFE489FB461246758B6B5Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Multicopy Overexpression of Bovine Pancreatic Trypsin Inhibitor Saturates the Protein Folding and Secretory Capacity of Saccharomyces cerevisiae</title><author><name sortKey="Parekh, R" sort="Parekh, R" uniqKey="Parekh R" first="R." last="Parekh">R. Parekh</name></author><author><name sortKey="Forrester, K" sort="Forrester, K" uniqKey="Forrester K" first="K." last="Forrester">K. Forrester</name></author><author><name sortKey="Wittrup, D" sort="Wittrup, D" uniqKey="Wittrup D" first="D." last="Wittrup">D. Wittrup</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:3EC9F7D7DAC6F7402AEDFE489FB461246758B6B5</idno><date when="1995" year="1995">1995</date><idno type="doi">10.1006/prep.1995.1071</idno><idno type="url">https://api.istex.fr/ark:/67375/6H6-VMRQ891Q-9/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">001606</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">001606</idno><idno type="wicri:Area/Istex/Curation">001606</idno><idno type="wicri:Area/Istex/Checkpoint">001843</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">001843</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Multicopy Overexpression of Bovine Pancreatic Trypsin Inhibitor Saturates the Protein Folding and Secretory Capacity of Saccharomyces cerevisiae</title><author><name sortKey="Parekh, R" sort="Parekh, R" uniqKey="Parekh R" first="R." last="Parekh">R. Parekh</name><affiliation wicri:level="1"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Univ Illinois, Dept Chem Engn, Urbana, IL 61801</wicri:regionArea><wicri:noRegion>IL 61801</wicri:noRegion></affiliation></author><author><name sortKey="Forrester, K" sort="Forrester, K" uniqKey="Forrester K" first="K." last="Forrester">K. Forrester</name><affiliation wicri:level="1"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Univ Illinois, Dept Chem Engn, Urbana, IL 61801</wicri:regionArea><wicri:noRegion>IL 61801</wicri:noRegion></affiliation></author><author><name sortKey="Wittrup, D" sort="Wittrup, D" uniqKey="Wittrup D" first="D." last="Wittrup">D. Wittrup</name><affiliation wicri:level="1"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Univ Illinois, Dept Chem Engn, Urbana, IL 61801</wicri:regionArea><wicri:noRegion>IL 61801</wicri:noRegion></affiliation></author></analytic><monogr></monogr><series><title level="j">Protein Expression and Purification</title><title level="j" type="abbrev">YPREP</title><idno type="ISSN">1046-5928</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1995">1995</date><biblScope unit="volume">6</biblScope><biblScope unit="issue">4</biblScope><biblScope unit="page" from="537">537</biblScope><biblScope unit="page" to="545">545</biblScope></imprint><idno type="ISSN">1046-5928</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">1046-5928</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Bovine pancreatic trypsin inhibitor (BPTI) was expressed and secreted from a synthetic gene as a model system for the study of protein folding and secretion in Saccharomyces cerevisiae. The efficiency of different leader sequences in directing BPTI secretion was examined, and up to 11 μg/ml of active BPTI was secreted. In some fusion constructs, inefficient proteolytic processing by Kex2p, Ste13p, and signal peptidase were observed immediately adjacent to the BPTI N terminus. Insertion of dipeptide spacers improved endoproteolytic processing substantially but the level of secretion was unchanged, Overexpression from a 2-μm multicopy vector results in essentially unchanged BPTI secretion as compared to expression from a single copy centromere vector. BPTI expressed from a multicopy vector accumulates intracellularly in an unfolded form, indicating that available secretory chaperones and foldases can be saturated by increasing the rate of BPTI synthesis.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country></list><tree><country name="États-Unis"><noRegion><name sortKey="Parekh, R" sort="Parekh, R" uniqKey="Parekh R" first="R." last="Parekh">R. Parekh</name></noRegion><name sortKey="Forrester, K" sort="Forrester, K" uniqKey="Forrester K" first="K." last="Forrester">K. Forrester</name><name sortKey="Wittrup, D" sort="Wittrup, D" uniqKey="Wittrup D" first="D." last="Wittrup">D. Wittrup</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Istex/Checkpoint
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 001843 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Istex/Checkpoint/biblio.hfd -nk 001843 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= Istex
|étape= Checkpoint
|type= RBID
|clé= ISTEX:3EC9F7D7DAC6F7402AEDFE489FB461246758B6B5
|texte= Multicopy Overexpression of Bovine Pancreatic Trypsin Inhibitor Saturates the Protein Folding and Secretory Capacity of Saccharomyces cerevisiae
}}
| This area was generated with Dilib version V0.6.33. |  |