Protein Isoaspartate Methyltransferase Is a Multicopy Suppressor of Protein Aggregation in Escherichia coli

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dc.contributor.author Kern, Renee
dc.contributor.author Malki, Abderrahim
dc.contributor.author Abdallah, Jad
dc.contributor.author Lebart, Jean-Claude
dc.contributor.author Dubucs, Catherine
dc.contributor.author Hee Yu, Myeong
dc.contributor.author Richarme, Gilbert
dc.date.accessioned 2016-07-19T09:46:36Z
dc.date.available 2016-07-19T09:46:36Z
dc.date.copyright 2005 en_US
dc.date.issued 2016-07-19
dc.identifier.issn 0021-9193 en_US
dc.identifier.uri http://hdl.handle.net/10725/4167
dc.description.abstract We used preS2-S′-β-galactosidase, a three-domain fusion protein that aggregates extensively at 43°C in the cytoplasm of Escherichia coli, to search for multicopy suppressors of protein aggregation and inclusion body formation and took advantage of the known differential solubility of preS2-S′-β-galactosidase at 37 and 43°C to develop a selection procedure for the gene products that would prevent its aggregation in vivo at 43°C. First, we demonstrate that the differential solubility of preS2-S′-β-galactosidase results in a lactose-positive phenotype at 37°C as opposed to a lactose-negative phenotype at 43°C. We searched for multicopy suppressors of preS2-S′-β-galactosidase aggregation by selecting pink lactose-positive colonies on a background of white lactose-negative colonies at 43°C after transformation of bacteria with an E. coli gene bank. We discovered that protein isoaspartate methyltransferase (PIMT) is a multicopy suppressor of preS2-S′-β-galactosidase aggregation at 43°C. Overexpression of PIMT reduces the amount of preS2-S′-β-galactosidase found in inclusion bodies at 43°C and increases its amount in soluble fractions. It reduces the level of isoaspartate formation in preS2-S′-β-galactosidase and increases its thermal stability in E. coli crude extracts without increasing the thermostability of a control protein, citrate synthase, in the same extracts. We could not detect any induction of the heat shock response resulting from PIMT overexpression, as judged from amounts of DnaK and GroEL, which were similar in the PIMT-overproducing and control strains. These results suggest that PIMT might be overburdened in some physiological conditions and that its overproduction may be beneficial in conditions in which protein aggregation occurs, for example, during biotechnological protein overproduction or in protein aggregation diseases. en_US
dc.language.iso en en_US
dc.title Protein Isoaspartate Methyltransferase Is a Multicopy Suppressor of Protein Aggregation in Escherichia coli en_US
dc.type Article en_US
dc.description.version Published en_US
dc.author.school SOP en_US
dc.author.idnumber 200703820 en_US
dc.author.department Pharmaceutical Sciences Department en_US
dc.description.embargo N/A en_US
dc.relation.journal Journal of Bacteriology en_US
dc.journal.volume 187 en_US
dc.journal.issue 4 en_US
dc.article.pages 1377-1383 en_US
dc.identifier.doi http://dx.doi.org/10.1128/JB.187.4.1377-1383.2005 en_US
dc.identifier.ctation Kern, R., Malki, A., Abdallah, J., Liebart, J. C., Dubucs, C., Yu, M. H., & Richarme, G. (2005). Protein isoaspartate methyltransferase is a multicopy suppressor of protein aggregation in Escherichia coli. Journal of bacteriology, 187(4), 1377-1383. en_US
dc.author.email jabdallah@lau.edu.lb en_US
dc.identifier.tou http://libraries.lau.edu.lb/research/laur/terms-of-use/articles.php en_US
dc.identifier.url http://jb.asm.org/content/187/4/1377.short en_US
dc.orcid.id https://orcid.org/0000-0001-5267-4953 en_US

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