Marek_2020_Comput.Struct.Biotechnol.J_18_1352

Reference

Title : Structural and catalytic effects of surface loop-helix transplantation within haloalkane dehalogenase family - Marek_2020_Comput.Struct.Biotechnol.J_18_1352
Author(s) : Marek M , Chaloupkova R , Prudnikova T , Sato Y , Rezacova P , Nagata Y , Kuta Smatanova I , Damborsky J
Ref : Comput Struct Biotechnol J , 18 :1352 , 2020
Abstract :

Engineering enzyme catalytic properties is important for basic research as well as for biotechnological applications. We have previously shown that the reshaping of enzyme access tunnels via the deletion of a short surface loop element may yield a haloalkane dehalogenase variant with markedly modified substrate specificity and enantioselectivity. Here, we conversely probed the effects of surface loop-helix transplantation from one enzyme to another within the enzyme family of haloalkane dehalogenases. Precisely, we transplanted a nine-residue long extension of L9 loop and beta4 helix from DbjA into the corresponding site of DbeA. Biophysical characterization showed that this fragment transplantation did not affect the overall protein fold or oligomeric state, but lowered protein stability (DeltaT (m) = -5 to 6 degC). Interestingly, the crystal structure of DbeA mutant revealed the unique structural features of enzyme access tunnels, which are known determinants of catalytic properties for this enzyme family. Biochemical data confirmed that insertion increased activity of DbeA with various halogenated substrates and altered its enantioselectivity with several linear beta-bromoalkanes. Our findings support a protein engineering strategy employing surface loop-helix transplantation for construction of novel protein catalysts with modified catalytic properties.

PubMedSearch : Marek_2020_Comput.Struct.Biotechnol.J_18_1352
PubMedID: 32612758
Gene_locus related to this paper: brael-e2rv62

Related information

Gene_locus brael-e2rv62
Structure brael-e2rv62    6XY9

Citations formats

Marek M, Chaloupkova R, Prudnikova T, Sato Y, Rezacova P, Nagata Y, Kuta Smatanova I, Damborsky J (2020)
Structural and catalytic effects of surface loop-helix transplantation within haloalkane dehalogenase family
Comput Struct Biotechnol J 18 :1352

Marek M, Chaloupkova R, Prudnikova T, Sato Y, Rezacova P, Nagata Y, Kuta Smatanova I, Damborsky J (2020)
Comput Struct Biotechnol J 18 :1352