A kinetically stable plant subtilase with unique peptide mass fingerprints and dimerization properties

Yadav, Subhash Chandra; Jagannadham, Medicherla V.; Kundu, Suman

doi:10.1016/j.bpc.2008.09.019

Cited by 11 publications

(2 citation statements)

References 56 publications

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“…A tryptic digested sample of benghalensin was mixed with matrix and spotted on the polished stainless steel MALDI target plate for MALDI analysis. de novo sequences of peptides were determined by using the reported protocol (37).…”

Section: Methodsmentioning

confidence: 99%

Benghalensin, a Highly Stable Serine Protease from the Latex of Medicinal PlantFicus benghalensis

Sharma¹,

Kumari²,

Jagannadham³

2009

J. Agric. Food Chem.

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A serine protease was purified to homogeneity from the latex of medicinal plant Ficus benghalensis by a single step procedure using anion exchange chromatography. The enzyme, named benghalensin, has a molecular mass of 47 kDa (MALDI-TOF and SDS-PAGE). The purified protein is a glycoprotein, and the enzymatic activity is solely inhibited by PMSF and chymostatin, indicating that the enzyme belongs to the serine protease class. The isoelectric point of the enzyme is pH 4.4 with optimum pH and temperature of pH 8.0 and 55 degrees C respectively. The extinction coefficient (epsilon(1%)(280)) of the enzyme is 29.25, and the molecular structure consists of 17 tryptophan, 31 tyrosine and 09 cysteine residues. Peptide mass fingerprinting and de novo sequencing of tryptic-digested fragments of the protein did not find any putative conserved domains in BLAST analysis. The enzyme is stable and retains full activity over a broad range of pH and temperature or prolonged storage at 4 degrees C. Simple purification, high yield and stability enable exploration of the protein for structure-function relationship studies as well as other applications.

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Section: Methodsmentioning

confidence: 99%

Benghalensin, a Highly Stable Serine Protease from the Latex of Medicinal PlantFicus benghalensis

Sharma¹,

Kumari²,

Jagannadham³

2009

J. Agric. Food Chem.

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“…Our laboratory is involved in identifying the proteins from species of unknown genome sequences. 21–23 Earlier studies in our laboratory revealed that the relative intensity of b-ions increased for N-terminal acetylated peptides. 20,24 .…”

Section: Introductionmentioning

confidence: 97%

Characterization of acetylated histidine b₁-ion structure: A competition between oxazolone and side chain imidazole moiety

Yadav

Rao

Srinivas

et al. 2018

Eur J Mass Spectrom (Chichester)

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The detection of post-translational modifications of proteins is an important comprehensive research area. Over the years, proteomic studies involving protein acetylation have attracted a great deal of attention. In the present study, we have focussed on the acetylation of histidine and the intrinsic stability of b-ion of oxazolone ring and/or with side chain imidazole bicyclic product. The formation of oxazolone structure may occur when an amino moiety undergoes acetylation reaction and when it is present in the vicinity of the side chain imidazole moiety. Tryptic peptides generated from the proteins of Acenitobacter radioresistens MMC5-containing N-terminal histidine were explored in a standard proteomic workflow. Formation of [Formula: see text] ion with an oxazolone ring in these peptides has been supported by a tandem mass spectrometric study of a synthetic peptide and density functional theory calculations. The results obtained from this study have implications in understanding the fragmentation of the peptides generated in the proteomic workflows.

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Complete conformational stability of kinetically stable dimeric serine protease milin against pH, temperature, urea, and proteolysis

Yadav

Jagannadham

2009

Eur Biophys J

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Spectroscopic, calorimetric, and proteolytic methods were utilized to evaluate the stability of the kinetically stable, differentially glycosylated, dimeric serine protease milin as a function of pH (1.0-11.0), temperature, urea, and GuHCl denaturation in presence of 8 M urea at pH 2.0. The stability of milin remains equivalent to that of native at pH 1.0-11.0. However, negligible and reversible alteration in structure upon temperature transition has been observed at pH 2.0 and with 1.6 M GuHCl. Irreversible and incomplete calorimetric transition with apparent T (m) > 100 degrees C was observed at basic pH (9.0 and 10.0). Urea-induced unfolding at pH 4.0, and at pH 2.0 with GuHCl, in presence of 8 M urea also reveals incomplete unfolding. Milin has been found to exhibit proteolytic resistant in either native or denatured state against various commercial proteases. These results imply that the high conformational stability of milin against various denaturating conditions enable its potential use in protease-based industries.

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A kinetically stable plant subtilase with unique peptide mass fingerprints and dimerization properties

Cited by 11 publications

References 56 publications

Benghalensin, a Highly Stable Serine Protease from the Latex of Medicinal PlantFicus benghalensis

Benghalensin, a Highly Stable Serine Protease from the Latex of Medicinal PlantFicus benghalensis

Characterization of acetylated histidine b₁-ion structure: A competition between oxazolone and side chain imidazole moiety

Complete conformational stability of kinetically stable dimeric serine protease milin against pH, temperature, urea, and proteolysis

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A kinetically stable plant subtilase with unique peptide mass fingerprints and dimerization properties

Cited by 11 publications

References 56 publications

Benghalensin, a Highly Stable Serine Protease from the Latex of Medicinal PlantFicus benghalensis

Benghalensin, a Highly Stable Serine Protease from the Latex of Medicinal PlantFicus benghalensis

Characterization of acetylated histidine b1-ion structure: A competition between oxazolone and side chain imidazole moiety

Complete conformational stability of kinetically stable dimeric serine protease milin against pH, temperature, urea, and proteolysis

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Characterization of acetylated histidine b₁-ion structure: A competition between oxazolone and side chain imidazole moiety