Abstract:
BACKGROUND:Haloalkane dehalogenases (EC 3.8.1.5, HLDs) are α/β-hydrolases which catalyze the irreversible cleavage of carbon-halogen bonds of haloalkanes, producing an alcohol, a halide and a hydrogen ion. Haloalkanes are acutely toxic to animals and humans and their toxic effects are mainly observed in the liver, kidneys and central nervous system. OBJECTIVE:In the present work, the haloalkane dehalogenase from Rhizobium leguminosarum bv. trifolii (DrlA) was characterized. METHOD:Reverse transcription polymerase chain reaction analysis and enzyme activity assays revealed that the DrlA gene expression in R. leguminosarum bv. trifolii is induced by 1,2- dibromoethane (1,2-DBE) during the early exponential phase. The gene of the enzyme was isolated, cloned and expressed in E. coli Rosetta (DE3). RESULTS:Recombinant DrlA displays its high catalytic activity towards 1,2-DBE and the long-chain haloalkane 1-iodohexane. Limited activity was observed for other aliphatic and cyclic haloalkanes, indicating that the enzyme displays restricted substrate specificity, compared to other bacterial HLDs. Homology modelling and phylogenetic analysis suggested that the enzyme belongs to the HLD-II subfamily and shares the same overall fold and domain organization as other bacterial HLDs, however major variations were identified at the hydrophobic substrate-binding cavity, the cap domain and the entrance of the main tunnel that affect the size of the active site pocket and the substrate recognition mechanism. CONCLUSION:This work sheds new light on the environmental fate and toxicity of 1,2-DBE and provides new knowledge on the structure, function and diversity of HLDs for developing applications in toxicology.
journal_name
Protein Pept Lettjournal_title
Protein and peptide lettersauthors
Georgakis N,Chronopoulou E,Gad MA,Skliros D,Efrose R,Flemetakis E,Labrou NEdoi
10.2174/0929866524666170621094531subject
Has Abstractpub_date
2017-01-01 00:00:00pages
599-608issue
7eissn
0929-8665issn
1875-5305pii
PPL-EPUB-84285journal_volume
24pub_type
杂志文章abstract::Porins are integral membrane proteins found in the outer membrane of bacteria, mitochondria and chloroplasts. Herein, we have reviewed sequence and structural understanding about bacterial porins. The first porin structure from Rhodobacter capsulatus at 1.8 Å resolution in 1991 till the recent structural advancement, ...
journal_title:Protein and peptide letters
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更新日期:2015-01-01 00:00:00
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
pub_type: 杂志文章,评审
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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更新日期:2013-03-01 00:00:00
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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更新日期:2010-06-01 00:00:00
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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更新日期:2008-01-01 00:00:00
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journal_title:Protein and peptide letters
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更新日期:2007-01-01 00:00:00
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journal_title:Protein and peptide letters
pub_type: 杂志文章,评审
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更新日期:2019-01-01 00:00:00
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journal_title:Protein and peptide letters
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journal_title:Protein and peptide letters
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更新日期:2009-01-01 00:00:00
abstract:BACKGROUND:Laminin, a member of the Extracellular Matrix (ECM), is a glycoprotein that is used as a factor that affects cell adhesion, proliferation, survival, and differentiation. Of these, five globular domains (LG domains) of the alpha chain play an important role in influencing the cell by binding to the integrin. ...
journal_title:Protein and peptide letters
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更新日期:2019-01-01 00:00:00
abstract::The formation of local structure, in short peptides has been probed by examining cleavage patterns and rates of proteolysis of designed sequences with a high tendency to form beta-hairpin structures. Three model sequences which bear fluorescence donor and acceptor groups have been investigated: [see text]. Fluorescenc...
journal_title:Protein and peptide letters
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更新日期:2006-01-01 00:00:00