%0 Journal Article %1 ISI:000386866500003 %A Amm, Ingo %A Kawan, Mona %A Wolf, Dieter H. %C 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA %D 2016 %I ACADEMIC PRESS INC ELSEVIER SCIENCE %J ANALYTICAL BIOCHEMISTRY %K Chaperones; Cytosolic Luciferase; Proteasomal beta-isopropylmalate control; degradation; dehydrogenase} protein proteins; quality {Misfolded %P 14-21 %R 10.1016/j.ab.2016.09.012 %T Characterization of protein quality control components via dual reporter-containing misfolded cytosolic model substrates %V 515 %X Protein misfolding and protein aggregation are causes of severe diseases as neurodegenerative disorders, diabetes and cancer. Therefore, the cell has to constantly monitor the folding status of its proteome. Chaperones and components of the ubiquitin-proteasome system are key players in the cellular protein quality control process. In order to characterize components of the protein quality control system in a well-established model eukaryote - the yeast Saccharomyces cerevisiae - we established new cytosolic model substrates based on firefly luciferase and (beta-isopropylmalate dehydrogenase (Leu2). The use of these two different enzymes arranged in tandem as reporters enabled us to analyse the folding status and the degradation propensity of these new model substrates in yeast cells mutated in components of the cellular protein quality control system. The Hsp70 chaperone system known to be essential in the cellular protein quality control was chosen as a model for showing the high value of the luciferase-based model substrates in the characterization of components of the cytosolic protein quality control system in yeast. (C) 2016 Elsevier Inc. All rights reserved.

@article{ISI:000386866500003, abstract = {{Protein misfolding and protein aggregation are causes of severe diseases as neurodegenerative disorders, diabetes and cancer. Therefore, the cell has to constantly monitor the folding status of its proteome. Chaperones and components of the ubiquitin-proteasome system are key players in the cellular protein quality control process. In order to characterize components of the protein quality control system in a well-established model eukaryote - the yeast Saccharomyces cerevisiae - we established new cytosolic model substrates based on firefly luciferase and (beta-isopropylmalate dehydrogenase (Leu2). The use of these two different enzymes arranged in tandem as reporters enabled us to analyse the folding status and the degradation propensity of these new model substrates in yeast cells mutated in components of the cellular protein quality control system. The Hsp70 chaperone system known to be essential in the cellular protein quality control was chosen as a model for showing the high value of the luciferase-based model substrates in the characterization of components of the cytosolic protein quality control system in yeast. (C) 2016 Elsevier Inc. All rights reserved.}}, added-at = {2017-05-18T11:32:12.000+0200}, address = {{525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA}}, affiliation = {{Wolf, DH (Reprint Author), Univ Stuttgart, Inst Biochem, Pfaffenwaldring 55, D-70569 Stuttgart, Germany. Amm, I (Reprint Author), BZH Heidelberg, Neuenheimer Feld 328, D-69120 Heidelberg, Germany. Amm, Ingo; Kawan, Mona; Wolf, Dieter H., Univ Stuttgart, Inst Biochem, Pfaffenwaldring 55, D-70569 Stuttgart, Germany. Kawan, Mona, ZMBH Heidelberg, Neuenheimer Feld 282, Heidelberg, Germany.}}, author = {Amm, Ingo and Kawan, Mona and Wolf, Dieter H.}, author-email = {{ingo.amm@bzh.uni-heidelberg.de dieter.wolf@ibc.uni-stuttgart.de}}, biburl = {https://puma.ub.uni-stuttgart.de/bibtex/24b611096fc72e9455d5d9e3edbad250b/hermann}, doi = {{10.1016/j.ab.2016.09.012}}, eissn = {{1096-0309}}, funding-acknowledgement = {{Deutsche Forschungsgemeinschaft (DFG) {[}WO 210/18-2]}}, funding-text = {{We thank Avrom Caplan for providing the yeast expression plasmid pRS316-GAL1-LUC. This work was supported by the Deutsche Forschungsgemeinschaft (DFG), grant number WO 210/18-2 to D. H. Wolf.}}, interhash = {bc18e8c5c8c6310e4b0d379fc6f8c0c7}, intrahash = {4b611096fc72e9455d5d9e3edbad250b}, issn = {{0003-2697}}, journal = {{ANALYTICAL BIOCHEMISTRY}}, keywords = {Chaperones; Cytosolic Luciferase; Proteasomal beta-isopropylmalate control; degradation; dehydrogenase} protein proteins; quality {Misfolded}, keywords-plus = {{UBIQUITIN-PROTEASOME-SYSTEM; ER-ASSOCIATED DEGRADATION; SACCHAROMYCES-CEREVISIAE; FIREFLY LUCIFERASE; HEAT-SHOCK; YEAST; HSP70; STRESS; UBR1; RESPONSES}}, language = {{English}}, month = {{DEC 15}}, number-of-cited-references = {{41}}, pages = {{14-21}}, publisher = {{ACADEMIC PRESS INC ELSEVIER SCIENCE}}, research-areas = {{Biochemistry \& Molecular Biology; Chemistry}}, times-cited = {{0}}, timestamp = {2017-05-18T09:32:12.000+0200}, title = {{Characterization of protein quality control components via dual reporter-containing misfolded cytosolic model substrates}}, type = {{Article}}, volume = {{515}}, web-of-science-categories = {{Biochemical Research Methods; Biochemistry \& Molecular Biology; Chemistry, Analytical}}, year = {{2016}} }