date: 2020-12-24T11:29:44Z
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pdf:docinfo:title: Description of Transport Tunnel in HaloalkaneDehalogenase Variant LinB D147C+L177C fromSphingobium japonicum
xmp:CreatorTool: LaTeX with hyperref
Keywords: bacterial enzyme; haloalkane dehalogenase; mutant form; crystallization; tertiary structure; disulfide bond; protein engineering; molecular dynamics; access tunnel; substrate specificity
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subject: The activity of enzymes with active sites buried inside their protein core highly depends on the efficient transport of substrates and products between the active site and the bulk solvent. The engineering of access tunnels in order to increase or decrease catalytic activity and specificity in a rational way is a challenging task. Here, we describe a combined experimental and computational approach to characterize the structural basis of altered activity in the haloalkane dehalogenase LinB D147C+L177C variant. While the overall protein fold is similar to the wild type enzyme and the other LinB variants, the access tunnels have been altered by introduced cysteines that were expected to form a disulfide bond. Surprisingly, the mutations have allowed several conformations of the amino acid chain in their vicinity, interfering with the structural analysis of the mutant by X-ray crystallography. The duration required for the growing of protein crystals changed from days to 1.5 years by introducing the substitutions. The haloalkane dehalogenase LinB D147C+L177C variant crystal structure was solved to 1.15 Å resolution, characterized and deposited to Protein Data Bank under PDB ID 6s06.
dc:creator: Iuliia Iermak, Oksana Degtjarik, Petra Havlickova, Michal Kuty, Radka Chaloupkova, Jiri Damborsky, Tatyana Prudnikova and Ivana Kuta Smatanova
dcterms:created: 2020-12-24T11:24:39Z
Last-Modified: 2020-12-24T11:29:44Z
dcterms:modified: 2020-12-24T11:29:44Z
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title: Description of Transport Tunnel in HaloalkaneDehalogenase Variant LinB D147C+L177C fromSphingobium japonicum
Last-Save-Date: 2020-12-24T11:29:44Z
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pdf:docinfo:keywords: bacterial enzyme; haloalkane dehalogenase; mutant form; crystallization; tertiary structure; disulfide bond; protein engineering; molecular dynamics; access tunnel; substrate specificity
pdf:docinfo:modified: 2020-12-24T11:29:44Z
meta:save-date: 2020-12-24T11:29:44Z
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dc:title: Description of Transport Tunnel in HaloalkaneDehalogenase Variant LinB D147C+L177C fromSphingobium japonicum
modified: 2020-12-24T11:29:44Z
cp:subject: The activity of enzymes with active sites buried inside their protein core highly depends on the efficient transport of substrates and products between the active site and the bulk solvent. The engineering of access tunnels in order to increase or decrease catalytic activity and specificity in a rational way is a challenging task. Here, we describe a combined experimental and computational approach to characterize the structural basis of altered activity in the haloalkane dehalogenase LinB D147C+L177C variant. While the overall protein fold is similar to the wild type enzyme and the other LinB variants, the access tunnels have been altered by introduced cysteines that were expected to form a disulfide bond. Surprisingly, the mutations have allowed several conformations of the amino acid chain in their vicinity, interfering with the structural analysis of the mutant by X-ray crystallography. The duration required for the growing of protein crystals changed from days to 1.5 years by introducing the substitutions. The haloalkane dehalogenase LinB D147C+L177C variant crystal structure was solved to 1.15 Å resolution, characterized and deposited to Protein Data Bank under PDB ID 6s06.
pdf:docinfo:subject: The activity of enzymes with active sites buried inside their protein core highly depends on the efficient transport of substrates and products between the active site and the bulk solvent. The engineering of access tunnels in order to increase or decrease catalytic activity and specificity in a rational way is a challenging task. Here, we describe a combined experimental and computational approach to characterize the structural basis of altered activity in the haloalkane dehalogenase LinB D147C+L177C variant. While the overall protein fold is similar to the wild type enzyme and the other LinB variants, the access tunnels have been altered by introduced cysteines that were expected to form a disulfide bond. Surprisingly, the mutations have allowed several conformations of the amino acid chain in their vicinity, interfering with the structural analysis of the mutant by X-ray crystallography. The duration required for the growing of protein crystals changed from days to 1.5 years by introducing the substitutions. The haloalkane dehalogenase LinB D147C+L177C variant crystal structure was solved to 1.15 Å resolution, characterized and deposited to Protein Data Bank under PDB ID 6s06.
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pdf:docinfo:creator: Iuliia Iermak, Oksana Degtjarik, Petra Havlickova, Michal Kuty, Radka Chaloupkova, Jiri Damborsky, Tatyana Prudnikova and Ivana Kuta Smatanova
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creator: Iuliia Iermak, Oksana Degtjarik, Petra Havlickova, Michal Kuty, Radka Chaloupkova, Jiri Damborsky, Tatyana Prudnikova and Ivana Kuta Smatanova
meta:author: Iuliia Iermak, Oksana Degtjarik, Petra Havlickova, Michal Kuty, Radka Chaloupkova, Jiri Damborsky, Tatyana Prudnikova and Ivana Kuta Smatanova
dc:subject: bacterial enzyme; haloalkane dehalogenase; mutant form; crystallization; tertiary structure; disulfide bond; protein engineering; molecular dynamics; access tunnel; substrate specificity
meta:creation-date: 2020-12-24T11:24:39Z
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meta:keyword: bacterial enzyme; haloalkane dehalogenase; mutant form; crystallization; tertiary structure; disulfide bond; protein engineering; molecular dynamics; access tunnel; substrate specificity
Author: Iuliia Iermak, Oksana Degtjarik, Petra Havlickova, Michal Kuty, Radka Chaloupkova, Jiri Damborsky, Tatyana Prudnikova and Ivana Kuta Smatanova
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