The production of soluble and correctly folded recombinant bovine β-lactoglobulin variants A and B in Escherichia coli for NMR studies

Ponniah, Komala; Loo, Trevor S.; Edwards, Patrick J. B.; Pascal, Steven M.; Jameson, Geoffrey B.; Norris, Gillian E.

doi:10.1016/j.pep.2009.12.006

Cited by 36 publications

(24 citation statements)

References 43 publications

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“…In unfavorable situations for disulfide bonds, configuration of the tertiary structure can cause damage and misfolding of the protein in E. coli. Such problems were explained for bovine β-lactoglobulin having five cysteine residues [12] and also for Nogo-A-specific exon 3 that has eight cysteine residues [6]. Export from the plasma membrane into the periclinal cell wall engages transfer of the comparatively unpolar scent molecules from a lipophilic into an aqueous section.…”

Section: Resultsmentioning

confidence: 99%

“…Under aerobic conditions; phenylacetaldehyde synthase (PAAS) catalyze oxidative decarboxylation by a radical mechanism. PAAS is the first PLP enzyme to be described that, in its native state, catalyzes the stoichiometric oxidative decarboxylation of an L-amino acid substrate ( Figure 1) [1][2][3][4][5][6][7][8][9][10][11][12][13]. PAAS by alignment is homologous with Aromatic L-amino acid decarboxylase (AADC) catalyzes the second enzymatic step in synthesis of the neurotransmitters dopamine and serotonin, which are found in neurons of all animals [4].…”

Section: Introductionmentioning

confidence: 99%

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Bioinformatics Analysis of Phenylacetaldehyde Synthase (PAAS), a Protein Involved in Flower Scent Production, in Rose

Jandoust¹

2014

J Phys Chem Biophys

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Rose flowers produce and emit the aromatic volatiles 2-phenylacetaldehyde (PAA) and 2-phenylethanol (2-PE), which have a distinctive flowery/rose-like scent. Previous studies of rose have shown that, similar to Petunia flowers, PAA is formed from L-phenylalanine via pyridoxal-5'-phosphate-dependent L-aromatic amino acid decarboxylase. Rosa phenylacetaldehyde synthase sequence (RhPAAS) is homologous to Petunia phenylacetaldehyde synthase (PhPAAS). Since there is not much experimental data available about different structural properties of that PAAS protein, in the present investigation, we studied the different structural properties of the PAAS protein in petunia and rose using bioinformatics tools. The features of the first, secondary and tertiary structures of this protein were compared between Petunia and Rose. The results indicated that the frequency of negatively charged, Leucine, and frequency of the Ser-Leu, Pro-Glu , Phe-Ser and the, Thr-Thr dipeptides in petunia are more than those in Rose. In contrast, in petunia, the frequencies of hydrophobic and hydrophilic residues, α-helix, β-sheet, β-strands of petunia are lower than those in rose. The features achieved in this study may also provide useful clues for designing scent production pathways using protein engineering techniques.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Bioinformatics Analysis of Phenylacetaldehyde Synthase (PAAS), a Protein Involved in Flower Scent Production, in Rose

Jandoust¹

2014

J Phys Chem Biophys

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“…The open reading frame of the synthetic gene for cβlg was ligated into MCS2. The pETDuet‐DsbC‐cβlg construct was transformed into competent Origami (DE3) cells (Novagen), as previously established [5], and plated on LB plates containing 15 μg/mL kanamycin, 100 μg/mL ampicillin and 12.5 μg/mL tetracycline. A pre‐culture was grown with shaking overnight at 37 °C in LB containing the same concentrations of antibiotics.…”

Section: Methodsmentioning

confidence: 99%

“…The pellet was then suspended in a small volume of 50 mM Tris, 50 mM NaCl, pH 7.5 and the solubilised cβlg was subjected to size‐exclusion chromatography (GE Healthcare). Bβlg was also purified, as per Ponniah et al [5].…”

Section: Methodsmentioning

confidence: 99%

Ultra‐high resolution crystal structure of recombinant caprine β‐lactoglobulin

et al. 2014

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a b s t r a c t b-Lactoglobulin (blg) is the most abundant whey protein in the milks of ruminant animals. While bovine blg has been subjected to a vast array of studies, little is known about the caprine ortholog. We present an ultra-high resolution crystal structure of caprine blg complemented by analytical ultracentrifugation and small-angle X-ray scattering data. In both solution and crystalline states caprine blg is dimeric (K D < 5 lM); however, our data suggest a flexible quaternary arrangement of subunits within the dimer. These structural findings will provide insight into relationships among structural, processing, nutritional and immunological characteristics that distinguish cow's and goat's milk. Structured summary of protein interactions:cBlg and cBlg bind by cosedimentation in solution (View interaction) bBlg and bBlg bind by cosedimentation in solution (View interaction) cBlg and cBlg bind by X ray scattering (View interaction) cBlg and cBlg bind by X-ray crystallography (View interaction)

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“…Some of these include NMR studies (Ponniah et al 2010), immunological studies , and functional studies for screening ribonucleolytic activities of proteins (Raines et al 2006;Raines et al 2008). As such, the proteins expressed in this study are properly folded wild-type APE1…”

Section: Discussionmentioning

confidence: 99%

Investigating the effects of expressing APE1 human population variants in cellular systems.

Ma¹

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The production of soluble and correctly folded recombinant bovine β-lactoglobulin variants A and B in Escherichia coli for NMR studies

Cited by 36 publications

References 43 publications

Bioinformatics Analysis of Phenylacetaldehyde Synthase (PAAS), a Protein Involved in Flower Scent Production, in Rose

Bioinformatics Analysis of Phenylacetaldehyde Synthase (PAAS), a Protein Involved in Flower Scent Production, in Rose

Ultra‐high resolution crystal structure of recombinant caprine β‐lactoglobulin

Investigating the effects of expressing APE1 human population variants in cellular systems.

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