Controlling proteolytic degradation of the methionine enriched MB-1Trp protein

Protein design is currently used for the creation of new proteins with desirable traits, which include a superior nutritional value. One of the challenges of protein design in this area is to achieve the production of stable native-like proteins that resist the proteolytic pressure of the organism u...

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Autores principales:	Sasseville,Maxime, St-Louis,Catherine, Khoudi,Habib, Beauregard,Marc
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2004
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582004000300004
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spelling	oai:scielo:S0717-345820040003000042005-05-31Controlling proteolytic degradation of the methionine enriched MB-1Trp proteinSasseville,MaximeSt-Louis,CatherineKhoudi,HabibBeauregard,Marc Protein design is currently used for the creation of new proteins with desirable traits, which include a superior nutritional value. One of the challenges of protein design in this area is to achieve the production of stable native-like proteins that resist the proteolytic pressure of the organism used for its production (the bioreactor). We report here the identification of a specific peptide bond sensitive to E. coli proteolysis in the designer protein MB-1Trp. In an attempt to reduce proteolysis, we have created a MB-1TrpHis gene library in which the two amino acids surrounding the peptide bond, N44 and L45, were randomized using degenerated oligonucleotides. The initial characterization of MB-1TrpHis N44E/L45V and MB-1TrpHis N44E/L45M, 2 variants of the library that were more resistant than the parent protein, was performed in order to investigate the nature of the mutants' resistance. Our results suggest that the mutants behaved like MB-1Trp regarding folding and thermal stability, and that proteolytic resistance is due to the elimination of the protease recognition site.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.7 n.3 20042004-12-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582004000300004en
institution	Scielo Chile
collection	Scielo Chile
language	English
description	Protein design is currently used for the creation of new proteins with desirable traits, which include a superior nutritional value. One of the challenges of protein design in this area is to achieve the production of stable native-like proteins that resist the proteolytic pressure of the organism used for its production (the bioreactor). We report here the identification of a specific peptide bond sensitive to E. coli proteolysis in the designer protein MB-1Trp. In an attempt to reduce proteolysis, we have created a MB-1TrpHis gene library in which the two amino acids surrounding the peptide bond, N44 and L45, were randomized using degenerated oligonucleotides. The initial characterization of MB-1TrpHis N44E/L45V and MB-1TrpHis N44E/L45M, 2 variants of the library that were more resistant than the parent protein, was performed in order to investigate the nature of the mutants' resistance. Our results suggest that the mutants behaved like MB-1Trp regarding folding and thermal stability, and that proteolytic resistance is due to the elimination of the protease recognition site.
author	Sasseville,Maxime St-Louis,Catherine Khoudi,Habib Beauregard,Marc
spellingShingle	Sasseville,Maxime St-Louis,Catherine Khoudi,Habib Beauregard,Marc Controlling proteolytic degradation of the methionine enriched MB-1Trp protein
author_facet	Sasseville,Maxime St-Louis,Catherine Khoudi,Habib Beauregard,Marc
author_sort	Sasseville,Maxime
title	Controlling proteolytic degradation of the methionine enriched MB-1Trp protein
title_short	Controlling proteolytic degradation of the methionine enriched MB-1Trp protein
title_full	Controlling proteolytic degradation of the methionine enriched MB-1Trp protein
title_fullStr	Controlling proteolytic degradation of the methionine enriched MB-1Trp protein
title_full_unstemmed	Controlling proteolytic degradation of the methionine enriched MB-1Trp protein
title_sort	controlling proteolytic degradation of the methionine enriched mb-1trp protein
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2004
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582004000300004
work_keys_str_mv	AT sassevillemaxime controllingproteolyticdegradationofthemethionineenrichedmb1trpprotein AT stlouiscatherine controllingproteolyticdegradationofthemethionineenrichedmb1trpprotein AT khoudihabib controllingproteolyticdegradationofthemethionineenrichedmb1trpprotein AT beauregardmarc controllingproteolyticdegradationofthemethionineenrichedmb1trpprotein
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Controlling proteolytic degradation of the methionine enriched MB-1Trp protein

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