Post‐translational modification and extended glycosylation pattern of a plant latex peroxidase of native source characterized by X‐ray crystallography

Abstract

The crystal structure of banyan peroxidase purified from the latex of Ficus benghalensis has been solved at 1.67 Å resolution by single‐wavelength anomalous diffraction phasing. The refined structure includes 306 amino acid residues, a heme and two calcium ions. The protein belongs to class III peroxidases and is the first one from plant latex. Extensive glycosylation was observed with N‐linked glycans attached to seven asparagine residues. The enzyme is stable with respect to a wide pH range, temperature, chemical denaturants and organic solvents, probably as a result of its high glycosylation. An unexpected post‐translational modification of Asp290 was identified as succinimide moiety. Kinetic parameters of banyan peroxidase have been determined using various hydrogen donor substrates and hydrogen peroxide. Database Coordinates and structure factors have been deposited in the Protein Data Bank under accession number http://www.rcsb.org/pdb/search/structidSearch.do?structureId=4CUO.

Arabidopsis thaliana root cell wall proteomics: Increasing the proteome coverage using a combinatorial peptide ligand library and description of unexpected Hyp in peroxidase amino acid sequences

“…On the contrary, Pro/Hyp residues of motif 3 were located within the substrate binding site, at the edge of the groove where the heme is bound (Fig. B) . The other observed Hyp residues in less conserved motifs were located at the surface of the R. sativus CIII Prx (data not shown).…”

“…Four such structures have been determined by X‐ray crystallography for native CIII Prxs bearing N ‐glycans, i.e. from peanut , royal palm tree , banyan tree , and Raphanus sativus (Gil‐Rodriguez et al., unpublished data). We have chosen that of R. sativus because this plant is a Brassicaceae like A. thaliana (PDB code 4A5G, http://www.rcsb.org/pdb/home/home.do).…”

Unexpected functional implication of a stable succinimide in the structural stability of Methanocaldococcus jannaschii glutaminase

“…Two recent examples of the succinimidyl group in the crystal structure of proteins include Thermus thermophilus amylomaltase and Ficus benghalensis peroxidase but neither the spontaneity of formation, nor the functional significance of this modification has been examined 51 52 . The succinimide in MjGATase is fully retained even after long periods of storage, boiling and prolonged incubation in high concentrations of chaotrope, strongly suggesting that this modification is remarkably stable in this enzyme.…”

How a novel tyrosine–heme cross-link fine-tunes the structure and functions of heme proteins: a direct comparitive study of L29H/F43Y myoglobin

“…myoglobin, Mb and hemoglobin, Hb), electron transfer (e.g. Examples include nitration of Tyr/Trp, [13][14][15] glycosylation of Asn, 16 and formation of cross-links, not only between amino acids such as Tyr-His in heme-copper oxidase (HCO), 17,18 but also between the heme and the protein matrix. peroxidase and cytochrome P450, CYP450), and signaling (NO sensor soluble guanylate cyclase, sGC, and CO sensor CooA, etc).…”