Research Interests:
Research Interests: NMR Spectroscopy, Crystallization, Sequence Analysis, Cell Culture, Biological Sciences, and 12 moreProtein-Protein Interaction, Circular Dichroism, Escherichia coli, Physical sciences, Structure Analysis, Protein Sequence Analysis, Spectrum, Secondary Structure, Amino Acid Profile, Aqueous Solution, Size Exclusion Chromatography, and Full Length Movies
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Research Interests:
Research Interests: Immunology, Small angle X-ray and neutron scattering, Magnetic Resonance Spectroscopy, Macromolecular X-Ray Crystallography, Crystal structure, and 13 moreMolecular Immunology, Humans, Staphylococcus aureus, Amino Acids, Protein Secondary Structure Prediction, Surface Properties, Amino Acid Sequence, Structure activity Relationship, Protein Binding, Ligands, Binding Site, Titrimetry, and Molecular Sequence Data
Research Interests:
Research Interests: Genetics, Symbiosis, Lepidoptera, Biological Sciences, Virulence, and 16 moreProtein Design, Humans, Bacillus thuringiensis, Bacterial Toxins, Animals, Temperature, Surface plasmon resonance, Temperature Dependence, Gram-negative bacteria, Nematoda, Galleria Mellonella, Two-Dimensional Gel Electrophoresis, Proteome, Bacterial Adhesion, Pathogenic Bacteria, and Photorhabdus
Members of the 2-oxoacid dehydrogenase multienzyme complex family play a key role in the pathways of central metabolism. Post-translational lipoylation of the dihydrolipoyl acyltransferase component of these complexes is essential for... more
Members of the 2-oxoacid dehydrogenase multienzyme complex family play a key role in the pathways of central metabolism. Post-translational lipoylation of the dihydrolipoyl acyltransferase component of these complexes is essential for their activity, the lipoyllysine moiety performing the transfer of substrates and intermediates between the different active sites within these multienzyme systems. We have previously shown that the thermophilic archaeon, Thermoplasma acidophilum, has a four-gene cluster encoding the components of such a complex, which, when recombinantly expressed in Escherichia coli, can be assembled into an active multienzyme in vitro. Crucially, the E. coli host carries out the required lipoylation of the archaeal dihydrolipoyl acyltransferase component. Because active 2-oxoacid dehydrogenase multienzyme complexes have never been detected in any archaeon, the question arises as to whether Archaea possess a functional lipoylation system. In this study, we report the cloning and heterologous expression of two genes from Tp. acidophilum whose protein products together show significant sequence identity with the single lipoate protein ligase enzyme of bacteria. We demonstrate that both recombinantly expressed Tp. acidophilum proteins are required for lipoylation of the acyltransferase, and that the two proteins associate together to carry out this post-translational modification. From the published DNA sequences, we suggest the presence of functional transcriptional and translational regulatory elements, and furthermore we present preliminary evidence that lipoylation occurs in vivo in Tp. acidophilum. This is the first report of the lipoylation machinery in the Archaea, which is unique in that the catalytic activity is dependent on two separate gene products.Structured digital abstract• MINT-7103712: E2lipD (uniprotkb:Q9HIA5), CTD (uniprotkb:Q9HKT2) and LplA (uniprotkb:Q9HKT1) physically interact (MI:0915) by molecular sieving (MI:0071)MINT-7103712: E2lipD (uniprotkb:Q9HIA5), CTD (uniprotkb:Q9HKT2) and LplA (uniprotkb:Q9HKT1) physically interact (MI:0915) by molecular sieving (MI:0071)
Research Interests:
Research Interests:
Research Interests:
Research Interests:
Research Interests: Psychology, Mathematics, Physics, Electronics, NMR Spectroscopy, and 20 moreMagnetic Resonance Spectroscopy, Body Language, Biological Sciences, Sequence alignment, Differential scanning calorimetry, Physical sciences, Travelling, Salmonella, Secondary Structure, Protein Secondary Structure Prediction, Analytical Ultracentrifugation, CD Spectroscopy, Amino Acid Profile, Amino Acid Sequence, Aqueous Solution, Hydrogen-Ion Concentration, Size Exclusion Chromatography, Rho GTPase, Functional Form, and Molecular Sequence Data
Research Interests:
Research Interests: NMR Spectroscopy, Crystallization, Sequence Analysis, Cell Culture, Biological Sciences, and 12 moreProtein-Protein Interaction, Circular Dichroism, Escherichia coli, Physical sciences, Structure Analysis, Protein Sequence Analysis, Spectrum, Secondary Structure, Amino Acid Profile, Aqueous Solution, Size Exclusion Chromatography, and Full Length Movies
Skip to content. University of Bath Opus: Online Publications Store. ...
Research Interests:
Research Interests: Immunology, Small angle X-ray and neutron scattering, Magnetic Resonance Spectroscopy, Macromolecular X-Ray Crystallography, Crystal structure, and 13 moreMolecular Immunology, Humans, Staphylococcus aureus, Amino Acids, Protein Secondary Structure Prediction, Surface Properties, Amino Acid Sequence, Structure activity Relationship, Protein Binding, Ligands, Binding Site, Titrimetry, and Molecular Sequence Data
Research Interests:
Research Interests: Genetics, Symbiosis, Lepidoptera, Biological Sciences, Virulence, and 16 moreProtein Design, Humans, Bacillus thuringiensis, Bacterial Toxins, Animals, Temperature, Surface plasmon resonance, Temperature Dependence, Gram-negative bacteria, Nematoda, Galleria Mellonella, Two-Dimensional Gel Electrophoresis, Proteome, Bacterial Adhesion, Pathogenic Bacteria, and Photorhabdus
Members of the 2-oxoacid dehydrogenase multienzyme complex family play a key role in the pathways of central metabolism. Post-translational lipoylation of the dihydrolipoyl acyltransferase component of these complexes is essential for... more
Members of the 2-oxoacid dehydrogenase multienzyme complex family play a key role in the pathways of central metabolism. Post-translational lipoylation of the dihydrolipoyl acyltransferase component of these complexes is essential for their activity, the lipoyllysine moiety performing the transfer of substrates and intermediates between the different active sites within these multienzyme systems. We have previously shown that the thermophilic archaeon, Thermoplasma acidophilum, has a four-gene cluster encoding the components of such a complex, which, when recombinantly expressed in Escherichia coli, can be assembled into an active multienzyme in vitro. Crucially, the E. coli host carries out the required lipoylation of the archaeal dihydrolipoyl acyltransferase component. Because active 2-oxoacid dehydrogenase multienzyme complexes have never been detected in any archaeon, the question arises as to whether Archaea possess a functional lipoylation system. In this study, we report the cloning and heterologous expression of two genes from Tp. acidophilum whose protein products together show significant sequence identity with the single lipoate protein ligase enzyme of bacteria. We demonstrate that both recombinantly expressed Tp. acidophilum proteins are required for lipoylation of the acyltransferase, and that the two proteins associate together to carry out this post-translational modification. From the published DNA sequences, we suggest the presence of functional transcriptional and translational regulatory elements, and furthermore we present preliminary evidence that lipoylation occurs in vivo in Tp. acidophilum. This is the first report of the lipoylation machinery in the Archaea, which is unique in that the catalytic activity is dependent on two separate gene products.Structured digital abstract• MINT-7103712: E2lipD (uniprotkb:Q9HIA5), CTD (uniprotkb:Q9HKT2) and LplA (uniprotkb:Q9HKT1) physically interact (MI:0915) by molecular sieving (MI:0071)MINT-7103712: E2lipD (uniprotkb:Q9HIA5), CTD (uniprotkb:Q9HKT2) and LplA (uniprotkb:Q9HKT1) physically interact (MI:0915) by molecular sieving (MI:0071)
