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Amino acid sequence of anionic peroxidase from the windmill palm tree Trachycarpus fortunei.

Baker MR, Zhao H, Sakharov IY, Li QX - J. Agric. Food Chem. (2014)

Bottom Line: Mature WPTP was 306 amino acids in length, and its carbohydrate content ranged from 21% to 29%.Comparison to closely related royal palm tree peroxidase revealed structural features that may explain differences in their substrate specificity.The results can be used to guide engineering of WPTP and its novel applications.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa , Honolulu, Hawaii 96822, United States.

ABSTRACT
Palm peroxidases are extremely stable and have uncommon substrate specificity. This study was designed to fill in the knowledge gap about the structures of a peroxidase from the windmill palm tree Trachycarpus fortunei. The complete amino acid sequence and partial glycosylation were determined by MALDI-top-down sequencing of native windmill palm tree peroxidase (WPTP), MALDI-TOF/TOF MS/MS of WPTP tryptic peptides, and cDNA sequencing. The propeptide of WPTP contained N- and C-terminal signal sequences which contained 21 and 17 amino acid residues, respectively. Mature WPTP was 306 amino acids in length, and its carbohydrate content ranged from 21% to 29%. Comparison to closely related royal palm tree peroxidase revealed structural features that may explain differences in their substrate specificity. The results can be used to guide engineering of WPTP and its novel applications.

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Alignment of WPTP and RPTP (PDB: 3HDL) according to themethod of Armougomet al.27 to determine features relatedto substrate specificity. Determination of (highlighted in gray) residuesless than 12 Å from heme iron and (highlighted in black) lessthan 4.6 Å from MES with Swiss-PdbViewer 4.1.028 and (box) residues with significant structural deviationbetween RPTP and HRP (PDB: 1ATJ) with a protein structure alignment tool.29 (&) Occupied glycosylation site;14 (c) cysteine; (+) Ca2+ binding. Secondarystructure: (dark gray) helices; (light gray) sheets. Helices labeledaccording to Watanabe et al.14 and conservationaccording to Livingston and Barton.30 Thefigure was drawn in JalView.31
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fig5: Alignment of WPTP and RPTP (PDB: 3HDL) according to themethod of Armougomet al.27 to determine features relatedto substrate specificity. Determination of (highlighted in gray) residuesless than 12 Å from heme iron and (highlighted in black) lessthan 4.6 Å from MES with Swiss-PdbViewer 4.1.028 and (box) residues with significant structural deviationbetween RPTP and HRP (PDB: 1ATJ) with a protein structure alignment tool.29 (&) Occupied glycosylation site;14 (c) cysteine; (+) Ca2+ binding. Secondarystructure: (dark gray) helices; (light gray) sheets. Helices labeledaccording to Watanabe et al.14 and conservationaccording to Livingston and Barton.30 Thefigure was drawn in JalView.31

Mentions: Class III peroxidases sharea common 3D structure despite a low sequence identity.2,16,26 For example, RPTP and HRP wereonly 36% identical, but their Cα backbone structures were closelyrelated, having an overall root mean squared deviation of 1.07 Å.14 A structurally guided alignment revealed thatWPTP and RPTP were 88% identical in primary structure and likely werevery similar in secondary and tertiary structures (Figure 5).


Amino acid sequence of anionic peroxidase from the windmill palm tree Trachycarpus fortunei.

Baker MR, Zhao H, Sakharov IY, Li QX - J. Agric. Food Chem. (2014)

Alignment of WPTP and RPTP (PDB: 3HDL) according to themethod of Armougomet al.27 to determine features relatedto substrate specificity. Determination of (highlighted in gray) residuesless than 12 Å from heme iron and (highlighted in black) lessthan 4.6 Å from MES with Swiss-PdbViewer 4.1.028 and (box) residues with significant structural deviationbetween RPTP and HRP (PDB: 1ATJ) with a protein structure alignment tool.29 (&) Occupied glycosylation site;14 (c) cysteine; (+) Ca2+ binding. Secondarystructure: (dark gray) helices; (light gray) sheets. Helices labeledaccording to Watanabe et al.14 and conservationaccording to Livingston and Barton.30 Thefigure was drawn in JalView.31
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4334278&req=5

fig5: Alignment of WPTP and RPTP (PDB: 3HDL) according to themethod of Armougomet al.27 to determine features relatedto substrate specificity. Determination of (highlighted in gray) residuesless than 12 Å from heme iron and (highlighted in black) lessthan 4.6 Å from MES with Swiss-PdbViewer 4.1.028 and (box) residues with significant structural deviationbetween RPTP and HRP (PDB: 1ATJ) with a protein structure alignment tool.29 (&) Occupied glycosylation site;14 (c) cysteine; (+) Ca2+ binding. Secondarystructure: (dark gray) helices; (light gray) sheets. Helices labeledaccording to Watanabe et al.14 and conservationaccording to Livingston and Barton.30 Thefigure was drawn in JalView.31
Mentions: Class III peroxidases sharea common 3D structure despite a low sequence identity.2,16,26 For example, RPTP and HRP wereonly 36% identical, but their Cα backbone structures were closelyrelated, having an overall root mean squared deviation of 1.07 Å.14 A structurally guided alignment revealed thatWPTP and RPTP were 88% identical in primary structure and likely werevery similar in secondary and tertiary structures (Figure 5).

Bottom Line: Mature WPTP was 306 amino acids in length, and its carbohydrate content ranged from 21% to 29%.Comparison to closely related royal palm tree peroxidase revealed structural features that may explain differences in their substrate specificity.The results can be used to guide engineering of WPTP and its novel applications.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa , Honolulu, Hawaii 96822, United States.

ABSTRACT
Palm peroxidases are extremely stable and have uncommon substrate specificity. This study was designed to fill in the knowledge gap about the structures of a peroxidase from the windmill palm tree Trachycarpus fortunei. The complete amino acid sequence and partial glycosylation were determined by MALDI-top-down sequencing of native windmill palm tree peroxidase (WPTP), MALDI-TOF/TOF MS/MS of WPTP tryptic peptides, and cDNA sequencing. The propeptide of WPTP contained N- and C-terminal signal sequences which contained 21 and 17 amino acid residues, respectively. Mature WPTP was 306 amino acids in length, and its carbohydrate content ranged from 21% to 29%. Comparison to closely related royal palm tree peroxidase revealed structural features that may explain differences in their substrate specificity. The results can be used to guide engineering of WPTP and its novel applications.

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