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  Description of Transport Tunnel in Haloalkane Dehalogenase Variant LinB D147C+L177C from Sphingobium japonicum

Iermak, I., Degtjarik, O., Havlickova, P., Kuty, M., Chaloupkova, R., Damborsky, J., et al. (2021). Description of Transport Tunnel in Haloalkane Dehalogenase Variant LinB D147C+L177C from Sphingobium japonicum. Catalysts, 11(1): 5. doi:10.3390/catal11010005.

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Item Permalink: https://hdl.handle.net/21.11116/0000-0007-F1AA-9 Version Permalink: https://hdl.handle.net/21.11116/0000-0007-F1AB-8
Genre: Journal Article

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catalysts-11-00005-v2.pdf (Publisher version), 11MB
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 Creators:
Iermak, Iuliia1, Author
Degtjarik, Oksana1, Author
Havlickova, Petra1, Author
Kuty, Michal1, Author
Chaloupkova, Radka1, Author
Damborsky, Jiri1, Author
Prudnikova, Tatyana1, Author
Kuta Smatanova, Ivana1, Author
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1external, ou_persistent22              

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Free keywords: Chemistry; bacterial enzyme; haloalkane dehalogenase; mutant form; crystallization; tertiary structure; disulfide bond; protein engineering; molecular dynamics; access tunnel; substrate specificity;
 Abstract: 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 angstrom resolution, characterized and deposited to Protein Data Bank under PDB ID 6s06.

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Language(s): eng - English
 Dates: 2021
 Publication Status: Published online
 Pages: 15
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: ISI: 000609997300001
DOI: 10.3390/catal11010005
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Title: Catalysts
  Abbreviation : Catalysts
Source Genre: Journal
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Publ. Info: Basel : MDPI
Pages: - Volume / Issue: 11 (1) Sequence Number: 5 Start / End Page: - Identifier: ISSN: 2073-4344
CoNE: https://pure.mpg.de/cone/journals/resource/2073-4344