Interplay Between Metal Binding and cis/trans Isomerization in Legume Lectins: Structural and Thermodynamic Study of P. angolensis Lectin

García-Pino, Abel; Buts, Lieven; Wyns, Lode; Loris, Remy

doi:10.1016/j.jmb.2006.06.006

Cited by 20 publications

(13 citation statements)

References 55 publications

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“…Mukwa lectin is much more thermostable than reported by Garcia-Pino et al [58], who determined the transition temperature from circular dichroism spectroscopy and differential scanning calorimetry to be about 76-78°C. The discrepancy between these results and those reported here can be explained by the presence of small amounts of Triton-X114 that were added by Garcia-Pino et al [58] to prevent precipitation of the protein at high temperature and promote reversibility of the process.…”

Section: Thermal Stabilitymentioning

confidence: 61%

“…Although the results of Garcia-Pino et al [58] point to monosaccharide (mannose) binding by the demetallized lectin, the latter apparently is not able anymore to accommodate larger oligosaccharides present on the glycoproteins ovalbumin and fetuin.…”

Section: Effect Of Demetallization On the Tertiary Structure And The mentioning

confidence: 91%

“…It was also observed that the conformation of D86 remains in cis upon demetallization because of the presence of the bulky H226 side chain that would clash with D86 upon cis-trans isomerization. Unexpectedly, the apo-lectin was also reported to remain able to bind mannose, though only very weakly [58].…”

Section: Effect Of Demetallization On the Tertiary Structure And The mentioning

confidence: 99%

“…Garcia-Pino et al [58] previously showed by X-ray crystallography that demetallization of mukwa lectin causes an important reorganization of the metal-binding loop whereby the Ca 2+ -binding site is completely destroyed. These authors also showed that the Mn 2+ -binding site is less affected by demetallization and that the primary sugar-binding site remains largely intact, though F132 and N138, two residues that are important for sugar-binding, are considerably displaced.…”

Section: Effect Of Demetallization On the Tertiary Structure And The mentioning

confidence: 99%

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Stability, Subunit Interactions and Carbohydrate-Binding of the Seed Lectin from Pterocarpus angolensis

Echemendia-Blanco

Driessche²,

Ncube³

et al. 2009

PPL

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From 1 kg of defatted Pterocarpus angolensis (mukwa tree) seed meal, 21.6 grams of an alpha,D-mannose/glucose-specific lectin can be purified on mannose-Sepharose. Relative affinities for several (oligo)saccharides and glycoproteins were studied by haemagglutination-inhibition. Gel filtration shows that the lectin exists as a dimer above pH 5 and as a monomer below pH 3.5. This is confirmed by studies on the release of lectin subunits that were adsorbed from solution to lectin monomers immobilized onto Eupergit-c. From the gel filtration patterns it is calculated that a residue with pK(a) of about 4.4 is involved in dimer dissociation. Titration of glutamic acids (E60, E209) is postulated to be involved. CD spectroscopy shows that the secondary structure of the lectin is unchanged between pH 1 and 12.5, and that the tertiary structure remains unchanged between pH 5 and 12. In the acid pH region, reversible spectral changes occur that may be due to the titration of one or more amino acids with a pK(a) value of 3.9-4.2, probably aspartic acid. These residues are implicated in sugar-binding but not in dimerization of the lectin. Only at pH 12.5, irreversible denaturation occurs. Mukwa lectin displays full carbohydrate-binding capacity between pH 4 and 12, as is concluded from ELLA (Enzyme Linked Lectin Assay) using ovalbumin and fetuin, and from binding of the same glycoproteins to immobilized lectin monomers. The lectin is rapidly and fully reversibly demetallized at pH 2.5 with 5 mM EDTA. The demetallized lectin is completely devoid of sugar-binding activity. Mukwa lectin is a very thermostable molecule (at least till 85 degrees C). However, addition of non-ionic detergents substantially lowers its thermostability.

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Section: Thermal Stabilitymentioning

confidence: 61%

Section: Effect Of Demetallization On the Tertiary Structure And The mentioning

confidence: 91%

Section: Effect Of Demetallization On the Tertiary Structure And The mentioning

confidence: 99%

Section: Effect Of Demetallization On the Tertiary Structure And The mentioning

confidence: 99%

See 2 more Smart Citations

Stability, Subunit Interactions and Carbohydrate-Binding of the Seed Lectin from Pterocarpus angolensis

Echemendia-Blanco

Driessche²,

Ncube³

et al. 2009

PPL

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show abstract

“…Like other L-type lectins, WFA possesses a conserved metal binding site for the divalent Ca 2ϩ and Mn 2ϩ ions. Their presence allow for the formation of a cis peptide bond between Ala-116 and Asp-117 (numbering scheme shown in Table 3), a common feature of legume lectins, which is essential for proper folding and carbohydrate binding (51).…”

Section: Resultsmentioning

confidence: 99%

Molecular Basis for Recognition of the Cancer Glycobiomarker, LacdiNAc (GalNAc[β1→4]GlcNAc), by Wisteria floribunda Agglutinin

Haji-Ghassemi

Gilbert

Spence

et al. 2016

Journal of Biological Chemistry

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Neoplastic transformation often results in cells with unusual glycosylation patterns specific to the type and stage of different cancers, providing numerous venues for therapeutic and diagnostic reagents (1, 2). Lectins are non-catalytic proteins with strict specificity for individual mono-or oligosaccharides, which can make them powerful tools for detecting changes in the carbohydrate structure in glycoproteins and glycolipids.

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Peptide Bond cis/trans Isomerases: A Biocatalysis Perspective of Conformational Dynamics in Proteins

Schiene‐Fischer

Aumüller

Fischer

2011

Topics in Current Chemistry

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Peptide bond cis/trans isomerases (PCTIases) catalyze an intrinsically slow rotational motion taking part in the conformational dynamics of a protein backbone in all of its folding states. In this way, PCTIases assist other proteins to shape their functionally active structure. They have been associated with viral, bacterial, and parasitic infection, signal transduction, cell differentiation, altered metabolic activity, apoptosis, and many other physiological and pathophysiological processes. The need to understand, characterize, and control biochemical steps which contribute to the folding of proteins is a problem being addressed in many laboratories today. This review discusses the biochemical basis that the peptidyl prolyl cis/trans isomerase (PPIase) family of PCTIases uses for the control of bioactivity. Special emphasis is given to recent developments in the field of biocatalytic features of PPIases, the mechanism of catalysis, and enzyme inhibition.

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Interplay Between Metal Binding and cis/trans Isomerization in Legume Lectins: Structural and Thermodynamic Study of P. angolensis Lectin

Cited by 20 publications

References 55 publications

Stability, Subunit Interactions and Carbohydrate-Binding of the Seed Lectin from Pterocarpus angolensis

Stability, Subunit Interactions and Carbohydrate-Binding of the Seed Lectin from Pterocarpus angolensis

Molecular Basis for Recognition of the Cancer Glycobiomarker, LacdiNAc (GalNAc[β1→4]GlcNAc), by Wisteria floribunda Agglutinin

Peptide Bond cis/trans Isomerases: A Biocatalysis Perspective of Conformational Dynamics in Proteins

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