Kuty M

References (14)

Title : Description of Transport Tunnel in Haloalkane Dehalogenase Variant LinB D147C+L177C from Sphingobium japonicum - Iermak_2021_Catalysts_11_5
Author(s) : Iermak I , Degtjarik O , Havlickova P , Kuty M , Chaloupkova R , Damborsky J , Prudnikova T , Smatanova IK
Ref : Catalysts , 11 :5 , 2021
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 A resolution, characterized and deposited to Protein Data Bank under PDB ID 6s06
ESTHER : Iermak_2021_Catalysts_11_5
PubMedSearch : Iermak_2021_Catalysts_11_5
PubMedID:
Gene_locus related to this paper: sphpi-linb

Title : The tetrameric structure of the novel haloalkane dehalogenase DpaA from Paraglaciecola agarilytica NO2 - Mazur_2021_Acta.Crystallogr.D.Struct.Biol_77_347
Author(s) : Mazur A , Prudnikova T , Grinkevich P , Mesters JR , Mrazova D , Chaloupkova R , Damborsky J , Kuty M , Kolenko P , Kuta Smatanova I
Ref : Acta Crystallographica D Struct Biol , 77 :347 , 2021
Abstract : Haloalkane dehalogenases (EC 3.8.1.5) are microbial enzymes that catalyse the hydrolytic conversion of halogenated compounds, resulting in a halide ion, a proton and an alcohol. These enzymes are used in industrial biocatalysis, bioremediation and biosensing of environmental pollutants or for molecular tagging in cell biology. The novel haloalkane dehalogenase DpaA described here was isolated from the psychrophilic and halophilic bacterium Paraglaciecola agarilytica NO2, which was found in marine sediment collected from the East Sea near Korea. Gel-filtration experiments and size-exclusion chromatography provided information about the dimeric composition of the enzyme in solution. The DpaA enzyme was crystallized using the sitting-drop vapour-diffusion method, yielding rod-like crystals that diffracted X-rays to 2.0A resolution. Diffraction data analysis revealed a case of merohedral twinning, and subsequent structure modelling and refinement resulted in a tetrameric model of DpaA, highlighting an uncommon multimeric nature for a protein belonging to haloalkane dehalogenase subfamily I.
ESTHER : Mazur_2021_Acta.Crystallogr.D.Struct.Biol_77_347
PubMedSearch : Mazur_2021_Acta.Crystallogr.D.Struct.Biol_77_347
PubMedID: 33645538
Gene_locus related to this paper: 9alte-k6xnl5

Title : Structural Analysis of the Ancestral Haloalkane Dehalogenase AncLinB-DmbA - Mazur_2021_Int.J.Mol.Sci_22_11992
Author(s) : Mazur A , Grinkevich P , Chaloupkova R , Havlickova P , Kascakova B , Kuty M , Damborsky J , Kuta Smatanova I , Prudnikova T
Ref : Int J Mol Sci , 22 : , 2021
Abstract : Haloalkane dehalogenases (EC 3.8.1.5) play an important role in hydrolytic degradation of halogenated compounds, resulting in a halide ion, a proton, and an alcohol. They are used in biocatalysis, bioremediation, and biosensing of environmental pollutants and also for molecular tagging in cell biology. The method of ancestral sequence reconstruction leads to prediction of sequences of ancestral enzymes allowing their experimental characterization. Based on the sequences of modern haloalkane dehalogenases from the subfamily II, the most common ancestor of thoroughly characterized enzymes LinB from Sphingobium japonicum UT26 and DmbA from Mycobacterium bovis 5033/66 was in silico predicted, recombinantly produced and structurally characterized. The ancestral enzyme AncLinB-DmbA was crystallized using the sitting-drop vapor-diffusion method, yielding rod-like crystals that diffracted X-rays to 1.5 A resolution. Structural comparison of AncLinB-DmbA with their closely related descendants LinB and DmbA revealed some differences in overall structure and tunnel architecture. Newly prepared AncLinB-DmbA has the highest active site cavity volume and the biggest entrance radius on the main tunnel in comparison to descendant enzymes. Ancestral sequence reconstruction is a powerful technique to study molecular evolution and design robust proteins for enzyme technologies.
ESTHER : Mazur_2021_Int.J.Mol.Sci_22_11992
PubMedSearch : Mazur_2021_Int.J.Mol.Sci_22_11992
PubMedID: 34769421
Gene_locus related to this paper: 9zzzz-AncLinB

Title : Stabilization of Haloalkane Dehalogenase Structure by Interfacial Interaction with Ionic Liquids - Shaposhnikova_2021_Crystals_11_1052
Author(s) : Shaposhnikova A , Kuty M , Chaloupkova R , Damborsky J , Smatanova IK , Minofar B , Prudnikova T
Ref : Crystals , 11 :1052 , 2021
Abstract : Ionic liquids attracted interest as green alternatives to replace conventional organic solvents in protein stability studies. They can play an important role in the stabilization of enzymes such as haloalkane dehalogenases that are used for biodegradation of warfare agents and halogenated environmental pollutants. Three-dimensional crystals of haloalkane dehalogenase variant DhaA80 (T148L+G171Q+A172V+C176F) from Rhodococcus rhodochrous NCIMB 13064 were grown and soaked with the solutions of 2-hydroxyethylammonium acetate and 1-butyl-3-methylimidazolium methyl sulfate. The objective was to study the structural basis of the interactions between the ionic liquids and the protein. The diffraction data were collected for the 1.25 A resolution for 2-hydroxyethylammonium acetate and 1.75 A resolution for 1-butyl-3-methylimidazolium methyl sulfate. The structures were used for molecular dynamics simulations to study the interactions of DhaA80 with the ionic liquids. The findings provide coherent evidence that ionic liquids strengthen both the secondary and tertiary protein structure due to extensive hydrogen bond interactions.
ESTHER : Shaposhnikova_2021_Crystals_11_1052
PubMedSearch : Shaposhnikova_2021_Crystals_11_1052
PubMedID:
Gene_locus related to this paper: rhoso-halo1

Title : Crystallization and Crystallographic Analysis of a Bradyrhizobium Elkanii USDA94 Haloalkane Dehalogenase Variant with an Eliminated Halide-Binding Site - Pudnikova_2019_Crystals_9_375
Author(s) : Prudnikova T , Kascakova B , Mesters JR , Grinkevich P , Havlickova P , Mazur A , Shaposhnikova A , Chaloupkova R , Damborsky J , Kuty M , Smatanova IK
Ref : , 9 :375 , 2019
Abstract : Haloalkane dehalogenases are a very important class of microbial enzymes for environmental detoxification of halogenated pollutants, for biocatalysis, biosensing and molecular tagging. The double mutant (Ile44Leu + Gln102His) of the haloalkane dehalogenase DbeA from Bradyrhizobium elkanii USDA94 (DbeADCl) was constructed to study the role of the second halide-binding site previously discovered in the wild-type structure. The variant is less active, less stable in the presence of chloride ions and exhibits significantly altered substrate specificity when compared with the DbeAwt. DbeADCl was crystallized using the sitting-drop vapour-diffusion procedure with further optimization by the random microseeding technique. The crystal structure of the DbeADCl has been determined and refined to the 1.4 A resolution. The DbeADCl crystals belong to monoclinic space group C121. The DbeADCl molecular structure was characterized and compared with five known haloalkane dehalogenases selected from the Protein Data Bank
ESTHER : Pudnikova_2019_Crystals_9_375
PubMedSearch : Pudnikova_2019_Crystals_9_375
PubMedID:
Gene_locus related to this paper: brael-e2rv62

Title : Crystal structure of the cold-adapted haloalkane dehalogenase DpcA from Psychrobacter cryohalolentis K5 - Tratsiak_2019_Acta.Crystallogr.F.Struct.Biol.Commun_75_324
Author(s) : Tratsiak K , Prudnikova T , Drienovska I , Damborsky J , Brynda J , Pachl P , Kuty M , Chaloupkova R , Rezacova P , Kuta Smatanova I
Ref : Acta Crystallographica F Struct Biol Commun , 75 :324 , 2019
Abstract : Haloalkane dehalogenases (HLDs) convert halogenated aliphatic pollutants to less toxic compounds by a hydrolytic mechanism. Owing to their broad substrate specificity and high enantioselectivity, haloalkane dehalogenases can function as biosensors to detect toxic compounds in the environment or can be used for the production of optically pure compounds. Here, the structural analysis of the haloalkane dehalogenase DpcA isolated from the psychrophilic bacterium Psychrobacter cryohalolentis K5 is presented at the atomic resolution of 1.05 A. This enzyme exhibits a low temperature optimum, making it attractive for environmental applications such as biosensing at the subsurface environment, where the temperature typically does not exceed 25 degrees C. The structure revealed that DpcA possesses the shortest access tunnel and one of the most widely open main tunnels among structural homologs of the HLD-I subfamily. Comparative analysis revealed major differences in the region of the alpha4 helix of the cap domain, which is one of the key determinants of the anatomy of the tunnels. The crystal structure of DpcA will contribute to better understanding of the structure-function relationships of cold-adapted enzymes.
ESTHER : Tratsiak_2019_Acta.Crystallogr.F.Struct.Biol.Commun_75_324
PubMedSearch : Tratsiak_2019_Acta.Crystallogr.F.Struct.Biol.Commun_75_324
PubMedID: 31045561
Gene_locus related to this paper: psyck-q1qbb9

Title : Crystallographic analysis of 1,2,3-trichloropropane biodegradation by the haloalkane dehalogenase DhaA31 - Lahoda_2014_Acta.Crystallogr.D.Biol.Crystallogr_70_209
Author(s) : Lahoda M , Mesters JR , Stsiapanava A , Chaloupkova R , Kuty M , Damborsky J , Kuta Smatanova I
Ref : Acta Crystallographica D Biol Crystallogr , 70 :209 , 2014
Abstract : Haloalkane dehalogenases catalyze the hydrolytic cleavage of carbon-halogen bonds, which is a key step in the aerobic mineralization of many environmental pollutants. One important pollutant is the toxic and anthropogenic compound 1,2,3-trichloropropane (TCP). Rational design was combined with saturation mutagenesis to obtain the haloalkane dehalogenase variant DhaA31, which displays an increased catalytic activity towards TCP. Here, the 1.31 A resolution crystal structure of substrate-free DhaA31, the 1.26 A resolution structure of DhaA31 in complex with TCP and the 1.95 A resolution structure of wild-type DhaA are reported. Crystals of the enzyme-substrate complex were successfully obtained by adding volatile TCP to the reservoir after crystallization at pH 6.5 and room temperature. Comparison of the substrate-free structure with that of the DhaA31 enzyme-substrate complex reveals that the nucleophilic Asp106 changes its conformation from an inactive to an active state during the catalytic cycle. The positions of three chloride ions found inside the active site of the enzyme indicate a possible pathway for halide release from the active site through the main tunnel. Comparison of the DhaA31 variant with wild-type DhaA revealed that the introduced substitutions reduce the volume and the solvent-accessibility of the active-site pocket.
ESTHER : Lahoda_2014_Acta.Crystallogr.D.Biol.Crystallogr_70_209
PubMedSearch : Lahoda_2014_Acta.Crystallogr.D.Biol.Crystallogr_70_209
PubMedID: 24531456
Gene_locus related to this paper: rhoso-halo1

Title : Structural and functional analysis of a novel haloalkane dehalogenase with two halide-binding sites - Chaloupkova_2014_Acta.Crystallogr.D.Biol.Crystallogr_70_1884
Author(s) : Chaloupkova R , Prudnikova T , Rezacova P , Prokop Z , Koudelakova T , Daniel L , Brezovsky J , Ikeda-Ohtsubo W4 , Sato Y , Kuty M , Nagata Y , Kuta Smatanova I , Damborsky J
Ref : Acta Crystallographica D Biol Crystallogr , 70 :1884 , 2014
Abstract : The crystal structure of the novel haloalkane dehalogenase DbeA from Bradyrhizobium elkanii USDA94 revealed the presence of two chloride ions buried in the protein interior. The first halide-binding site is involved in substrate binding and is present in all structurally characterized haloalkane dehalogenases. The second halide-binding site is unique to DbeA. To elucidate the role of the second halide-binding site in enzyme functionality, a two-point mutant lacking this site was constructed and characterized. These substitutions resulted in a shift in the substrate-specificity class and were accompanied by a decrease in enzyme activity, stability and the elimination of substrate inhibition. The changes in enzyme catalytic activity were attributed to deceleration of the rate-limiting hydrolytic step mediated by the lower basicity of the catalytic histidine.
ESTHER : Chaloupkova_2014_Acta.Crystallogr.D.Biol.Crystallogr_70_1884
PubMedSearch : Chaloupkova_2014_Acta.Crystallogr.D.Biol.Crystallogr_70_1884
PubMedID: 25004965
Gene_locus related to this paper: brael-e2rv62

Title : Differences in crystallization of two LinB variants from Sphingobium japonicum UT26 - Degtjarik_2013_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_69_284
Author(s) : Degtjarik O , Chaloupkova R , Rezacova P , Kuty M , Damborsky J , Kuta Smatanova I
Ref : Acta Crystallographica Sect F Struct Biol Cryst Commun , 69 :284 , 2013
Abstract : Haloalkane dehalogenases are microbial enzymes that convert a broad range of halogenated aliphatic compounds to their corresponding alcohols by the hydrolytic mechanism. These enzymes play an important role in the biodegradation of various environmental pollutants. Haloalkane dehalogenase LinB isolated from a soil bacterium Sphingobium japonicum UT26 has a relatively broad substrate specificity and can be applied in bioremediation and biosensing of environmental pollutants. The LinB variants presented here, LinB32 and LinB70, were constructed with the goal of studying the effect of mutations on enzyme functionality. In the case of LinB32 (L117W), the introduced mutation leads to blocking of the main tunnel connecting the deeply buried active site with the surrounding solvent. The other variant, LinB70 (L44I, H107Q), has the second halide-binding site in a position analogous to that in the related haloalkane dehalogenase DbeA from Bradyrhizobium elkanii USDA94. Both LinB variants were successfully crystallized and full data sets were collected for native enzymes as well as their complexes with the substrates 1,2-dibromoethane (LinB32) and 1-bromobutane (LinB70) to resolutions ranging from 1.6 to 2.8 A. The two mutants crystallize differently from each other, which suggests that the mutations, although deep inside the molecule, can still affect the protein crystallizability.
ESTHER : Degtjarik_2013_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_69_284
PubMedSearch : Degtjarik_2013_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_69_284
PubMedID: 23519805
Gene_locus related to this paper: sphpi-linb

Title : Crystallographic analysis of new psychrophilic haloalkane dehalogenases: DpcA from Psychrobacter cryohalolentis K5 and DmxA from Marinobacter sp. ELB17 - Tratsiak_2013_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_69_683
Author(s) : Tratsiak K , Degtjarik O , Drienovska I , Chrast L , Rezacova P , Kuty M , Chaloupkova R , Damborsky J , Kuta Smatanova I
Ref : Acta Crystallographica Sect F Struct Biol Cryst Commun , 69 :683 , 2013
Abstract : Haloalkane dehalogenases are hydrolytic enzymes with a broad range of potential practical applications such as biodegradation, biosensing, biocatalysis and cellular imaging. Two newly isolated psychrophilic haloalkane dehalogenases exhibiting interesting catalytic properties, DpcA from Psychrobacter cryohalolentis K5 and DmxA from Marinobacter sp. ELB17, were purified and used for crystallization experiments. After the optimization of crystallization conditions, crystals of diffraction quality were obtained. Diffraction data sets were collected for native enzymes and complexes with selected ligands such as 1-bromohexane and 1,2-dichloroethane to resolutions ranging from 1.05 to 2.49 A.
ESTHER : Tratsiak_2013_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_69_683
PubMedSearch : Tratsiak_2013_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_69_683
PubMedID: 23722854
Gene_locus related to this paper: 9alte-a3jb27 , psyck-q1qbb9

Title : Atomic resolution studies of haloalkane dehalogenases DhaA04, DhaA14 and DhaA15 with engineered access tunnels - Stsiapanava_2010_Acta.Crystallogr.D.Biol.Crystallogr_66_962
Author(s) : Stsiapanava A , Dohnalek J , Gavira JA , Kuty M , Koudelakova T , Damborsky J , Kuta Smatanova I
Ref : Acta Crystallographica D Biol Crystallogr , 66 :962 , 2010
Abstract : The haloalkane dehalogenase DhaA from Rhodococcus rhodochrous NCIMB 13064 is a bacterial enzyme that shows catalytic activity for the hydrolytic degradation of the highly toxic industrial pollutant 1,2,3-trichloropropane (TCP). Mutagenesis focused on the access tunnels of DhaA produced protein variants with significantly improved activity towards TCP. Three mutants of DhaA named DhaA04 (C176Y), DhaA14 (I135F) and DhaA15 (C176Y + I135F) were constructed in order to study the functional relevance of the tunnels connecting the buried active site of the protein with the surrounding solvent. All three protein variants were crystallized using the sitting-drop vapour-diffusion technique. The crystals of DhaA04 belonged to the orthorhombic space group P2(1)2(1)2(1), while the crystals of DhaA14 and DhaA15 had triclinic symmetry in space group P1. The crystal structures of DhaA04, DhaA14 and DhaA15 with ligands present in the active site were solved and refined using diffraction data to 1.23, 0.95 and 1.22 A, resolution, respectively. Structural comparisons of the wild type and the three mutants suggest that the tunnels play a key role in the processes of ligand exchange between the buried active site and the surrounding solvent.
ESTHER : Stsiapanava_2010_Acta.Crystallogr.D.Biol.Crystallogr_66_962
PubMedSearch : Stsiapanava_2010_Acta.Crystallogr.D.Biol.Crystallogr_66_962
PubMedID: 20823547
Gene_locus related to this paper: rhoso-halo1

Title : Pathways and mechanisms for product release in the engineered haloalkane dehalogenases explored using classical and random acceleration molecular dynamics simulations - Klvana_2009_J.Mol.Biol_392_1339
Author(s) : Klvana M , Pavlova M , Koudelakova T , Chaloupkova R , Dvorak P , Prokop Z , Stsiapanava A , Kuty M , Kuta-Smatanova I , Dohnalek J , Kulhanek P , Wade RC , Damborsky J
Ref : Journal of Molecular Biology , 392 :1339 , 2009
Abstract : Eight mutants of the DhaA haloalkane dehalogenase carrying mutations at the residues lining two tunnels, previously observed by protein X-ray crystallography, were constructed and biochemically characterized. The mutants showed distinct catalytic efficiencies with the halogenated substrate 1,2,3-trichloropropane. Release pathways for the two dehalogenation products, 2,3-dichloropropane-1-ol and the chloride ion, and exchange pathways for water molecules, were studied using classical and random acceleration molecular dynamics simulations. Five different pathways, denoted p1, p2a, p2b, p2c, and p3, were identified. The individual pathways showed differing selectivity for the products: the chloride ion releases solely through p1, whereas the alcohol releases through all five pathways. Water molecules play a crucial role for release of both products by breakage of their hydrogen-bonding interactions with the active-site residues and shielding the charged chloride ion during its passage through a hydrophobic tunnel. Exchange of the chloride ions, the alcohol product, and the waters between the buried active site and the bulk solvent can be realized by three different mechanisms: (i) passage through a permanent tunnel, (ii) passage through a transient tunnel, and (iii) migration through a protein matrix. We demonstrate that the accessibility of the pathways and the mechanisms of ligand exchange were modified by mutations. Insertion of bulky aromatic residues in the tunnel corresponding to pathway p1 leads to reduced accessibility to the ligands and a change in mechanism of opening from permanent to transient. We propose that engineering the accessibility of tunnels and the mechanisms of ligand exchange is a powerful strategy for modification of the functional properties of enzymes with buried active sites.
ESTHER : Klvana_2009_J.Mol.Biol_392_1339
PubMedSearch : Klvana_2009_J.Mol.Biol_392_1339
PubMedID: 19577578
Gene_locus related to this paper: rhoso-halo1

Title : Crystallization and preliminary X-ray analysis of a novel haloalkane dehalogenase DbeA from Bradyrhizobium elkani USDA94 - Prudnikova_2009_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_65_353
Author(s) : Prudnikova T , Mozga T , Rezacova P , Chaloupkova R , Sato Y , Nagata Y , Brynda J , Kuty M , Damborsky J , Smatanova IK
Ref : Acta Crystallographica Sect F Struct Biol Cryst Commun , 65 :353 , 2009
Abstract : A novel enzyme, DbeA, belonging to the haloalkane dehalogenase family (EC 3.8.1.5) was isolated from Bradyrhizobium elkani USDA94. This haloalkane dehalogenase is closely related to the DbjA enzyme from B. japonicum USDA110 (71% sequence identity), but has different biochemical properties. DbeA is generally less active and has a higher specificity towards brominated and iodinated compounds than DbjA. In order to understand the altered activity and specificity of DbeA, its mutant variant DbeA1, which carries the unique fragment of DbjA, was also constructed. Both wild-type DbeA and DbeA1 were crystallized using the sitting-drop vapour-diffusion method. The crystals of DbeA belonged to the primitive orthorhombic space group P2(1)2(1)2(1), while the crystals of DbeA1 belonged to the monoclinic space group C2. Diffraction data were collected to 2.2 A resolution for both DbeA and DbeA1 crystals.
ESTHER : Prudnikova_2009_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_65_353
PubMedSearch : Prudnikova_2009_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_65_353
PubMedID: 19342778
Gene_locus related to this paper: brael-e2rv62

Title : Computational site-directed mutagenesis of haloalkane dehalogenase in position 172 - Damborsky_1998_Protein.Eng_11_901
Author(s) : Damborsky J , Bohac M , Prokop M , Kuty M , Koca J
Ref : Protein Engineering , 11 :901 , 1998
Abstract : The application of molecular modelling and quantum-chemistry calculations for the 'computational site-directed mutagenesis' of haloalkane dehalogenase is described here. The exhaustive set of single point mutants of haloalkane dehalogenase in position 172 was constructed by homology modelling. The ability of substituting residues to stabilize the halide ion formed during the dehalogenation reaction in the enzyme active site was probed by quantum-chemical calculations. A simplified modelling procedure was adopted to obtain informative results on the potential activity of mutant proteins in a sufficiently short period of time, which, in the future, could be applicable for making bona fide predictions of mutants' activity prior to their preparation in the laboratory. The reaction pathways for the carbon-halide bond cleavage were calculated using microscopic models of wild type and mutant proteins. The theoretical parameters derived from the calculation, i.e. relative energies and selected atomic charges of educt, product and transition state structures, were statistically correlated with experimentally determined activities. The charge difference of educt and product on the halide-stabilizing hydrogen atom of residue 172 was the best parameter to distinguish protein variants with high activity from mutant proteins displaying a low activity. All mutants with significant activity in the experiment were found to have this parameter one order of magnitude higher than mutants with low activity. The results obtained are discussed in the light of the practical application of this methodology for the prediction of potentially active protein variants. Further automation of the modelling procedure is suggested for combinatorial screening of the large number of protein variants. Coupling of the dehalogenation reaction with hydrogenation of the halide ion formed during the reaction in the enzyme active site was proposed as a possible way to improve the catalytic activity of the haloalkane dehalogenase of Xanthobacter autotrophicus GJ10.
ESTHER : Damborsky_1998_Protein.Eng_11_901
PubMedSearch : Damborsky_1998_Protein.Eng_11_901
PubMedID: 9862209