Research article summary (published 15 Mar 2009):

First evidence for the salt-dependent folding and activity of an esterase from the halophilic archaea Haloarcula marismortui.

Full Abstract

A gene encoding an esterase from Haloarcula marismortui, a halophilic archaea from the Dead Sea, was cloned, expressed in Escherichia coli, and the recombinant protein (Hm EST) was biochemically characterized. The enzymatic activity of Hm EST was shown to exhibit salt dependence through salt-dependent folding. Hm EST exhibits a preference for short chain fatty acids and monoesters. It is inhibited by phenylmethylsulfonyl fluoride, diethyl-p-nitrophenyl phosphate, and 5-methoxy-3-(4-phenoxyphenyl)-3H-[1,3,4]oxadiazol-2-one, confirming the conclusion from sequence alignments that Hm EST is a serine carboxylesterase belonging to the hormone-sensitive lipase family. The activity of Hm EST is optimum in the presence of 3 M KCl and no activity was detected in the absence of salts. Far-UV circular dichroism showed that Hm EST is totally unfolded in salt-free medium and secondary structure appears in the presence of 0.25-0.5 M KCl. After salt depletion, the protein was able to recover 60% of its initial activity when 2 M KCl was added. A 3D model of Hm EST was built and its surface properties were analyzed, pointing to an enrichment in acidic residues paralleled by a depletion in basic residues. This peculiar charge repartition at the protein surface supports a better stability of the protein in a high salt environment.

Author information

Author/s: Müller-Santos, Marcelo (M); de Souza, Emanuel M (EM); Pedrosa, Fabio de O (Fde O); Mitchell, David Alexander (DA); Longhi, Sonia (S); Carrière, Frédéric (F); Canaan, Stéphane (S); Krieger, Nadia (N);

Affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Curitiba, Brazil.

Journal and publication information

Publication Type: Journal Article; Research Support, Non-U.S. Gov't

Journal: Biochimica et biophysica acta (Biochim Biophys Acta), published in Netherlands. (Language: eng)

Reference: 2009-Aug; vol 1791 (issue 8) : pp 719-29

Dates: Created 2009/07/20; Completed 2009/09/23;

PMID: 19303051, status: MEDLINE (last retrieved date: 9/23/2009)

Sourced from the National Library of Medicine. Abstract text and other information may be subject to copyright.

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MeSH headings (categories)

This article was linked to the MeSH Headings (categories) shown below.

Amino Acid Sequence Circular Dichroism Cloning, Molecular Computational Biology Electrophoresis, Polyacrylamide Gel Enzyme Activation - drug effects Enzyme Inhibitors - pharmacology Enzyme Stability - drug effects Esterases - chemistry; isolation & purification; metabolism Haloarcula marismortui - drug effects; enzymology Hydrogen-Ion Concentration Models, Molecular

Models, Molecular Molecular Sequence Data Potassium Chloride - pharmacology Protein Folding - drug effects Protein Structure, Secondary Sequence Alignment Sodium Chloride - pharmacology Static Electricity Substrate Specificity - drug effects Surface Properties - drug effects Temperature Time Factors

Note: Bold headings indicate primary MeSH headings or qualifiers.

Associated Chemicals: Enzyme Inhibitors (0) ; Potassium Chloride (7447-40-7) ; Sodium Chloride (7647-14-5) ; Esterases (EC 3.1.-)

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