C-terminal Post-translational Proteolysis of Plant Lectins and Their Recombinant Forms Expressed in Escherichia coli

Young, N. Martin; Watson, David; Yaguchi, Makoto; Adar, R; Arango, Rafael; Rodriguez-Arango, Esperanza; Sharon, Nathan; Blay, P. K. S.; Thibault, Pierre

doi:10.1074/jbc.270.6.2563

Cited by 51 publications

(40 citation statements)

References 33 publications

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“…The determinants were tentatively localized within a surface loop [118], but, because the results were obtained by fusion of truncated PHA with invertase, concerns about possible misfolding of the chimeric reporter protein would need to be addressed. The role of the C-terminal propeptide of PHA also requires further study, as it resembles the ctVSDs described above [122]. The theoretical role of a Cterminal propeptide in affecting folding and therefore presentation of hydrophobic patches has been discussed above and may be relevant for the following alternative model.…”

Section: The Physical Structure (Ps-vsd) Vacuolar Sorting Determinantsmentioning

confidence: 99%

Sorting of proteins to vacuoles in plant cells

Neuhaus

Rogers

1998

Protein Trafficking in Plant Cells

132

149

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An individual plant cell may contain at least two functionally and structurally distinct types of vacuoles: protein storage vacuoles and lytic vacuoles. Presumably a cell that stores proteins in vacuoles must maintain these separate compartments to prevent exposure of the storage proteins to an acidified environment with active hydrolytic enzymes where they would be degraded. Thus, the organization of the secretory pathway in plant cells, which includes the vacuoles, has a fascinating complexity not anticipated from the extensive genetic and biochemical studies of the secretory pathway in yeast. Plant cells must generate the membranes to form two separate types of tonoplast, maintain them as separate organelles, and direct soluble proteins from the secretory flow specifically to one or the other via separate vesicular pathways. Individual soluble and membrane proteins must be recognized and sorted into one or the other pathway by distinct, specific mechanisms. Here we review the emerging picture of how separate plant vacuoles are organized structurally and how proteins are recognized and sorted to each type.

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Section: The Physical Structure (Ps-vsd) Vacuolar Sorting Determinantsmentioning

confidence: 99%

Sorting of proteins to vacuoles in plant cells

Neuhaus

Rogers

1998

Protein Trafficking in Plant Cells

132

149

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“…Sato et al (38) also postulated that the C-terminal end is processed and cleaved between Asn-274 and Asn-275 by an enzyme, and our digest under reducing condition corroborated this finding with coverage up to Asn-274. Asn-specific cleavage of C-terminal peptides of legume lectins is a common phenomenon (59,60).…”

Section: Journal Of Biological Chemistry 24091mentioning

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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“…2). The appearance of double bands is most likely due to the C-terminal processing that legume lectins undergo both in the seeds and in the bacteria, as demonstrated by molecular weight measurements by electrospray mass spectrometry (Young et al, 1995).…”

Section: Physicochemical Characterizationmentioning

confidence: 99%

“…This result suggested that Tyr 108 plays a marginal role in the binding of the dansyl group by the lectin. Pro 134 appears not to be involved in ligand binding either, because, in the recombinant lectin, expressed in Escherichia coli, this residue is replaced by glutamine (Young et al, 1995), yielding approximately the same activity and affinity for different sugars as the native lectin (Arango et al, 1992(Arango et al, ,1993Adar & Sharon, 1996). To examine the role of Trp 135 and Tyr 108 in ligand binding, the former residue was replaced by tyrosine or alanine and the latter by threonine or alanine using site-directed mutagenesis.…”

Section: ) Although Their Quaternary Structures Are Sometimes DImentioning

confidence: 99%

“…After such automatic replacements, the side-chain conformations were adjusted, if necessary, taking into account rotamer populations and steric contacts with the rest of the protein. Each mutant listed in Table 1 carries an additional mutation because rECorL differs from ECorL at position 134 by having a Gln residue instead of Pro (Young et al, 1995). Finally, rECorL and the respective mutants were subjected to local energy minimization around the residue@) in question.…”

Section: Molecular Modelingmentioning

confidence: 99%

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Structural features of the combining site region of Erythrina corallodendron lectin: Role of tryptophan 135

et al. 1998

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The role of Trp 135 and Tyr 108 in the combining site of Erythrina corallodendron lectin (ECorL) was investigated by physicochemical characterization of mutants obtained by site‐directed mutagenesis, hemagglutination‐inhibition studies, and molecular modeling, including dynamics simulations. The findings demonstrate that Trp 135 in ECorL: (1) is required for the tight binding of Ca2+ and Mn2+ to the lectin because mutation of this residue into alanine results in loss of these ions upon dialysis and concomitant reversible inactivation of the mutant; (2) contributes to the high affinity of methyl α‐N‐dansylgalactosaminide (MeαGalNDns) to the lectin; and (3) is solely responsible for the fluorescence energy transfer between the aromatic residues of the lectin and the dansyl group in the ECorL‐MeαGalNDns complex. Docking of MeαGalNDns into the combining site of the lectin reveals that the dansyl moiety is parallel with the indole of Trp 135, as required for efficient fluorescence energy transfer, in one of the two possible conformations that this ligand assumes in the bound state. In the W135A mutant, which still binds MeαGalNDns strongly, the dansyl group may partially insert itself into the place formerly occupied by Trp 135, a process that from dynamics simulations does not appear to be energetically favored unless the loop containing this residue assumes an open conformation. However, a small fraction of the W135A molecules must be able to bind MeαGalNDns in order to explain the relatively high affinity, as compared to galactose, still remaining for this ligand. A model for the molecular events leading to inactivation of the W135A mutant upon demetallization is also presented in which the cis‐trans isomerization of the Ala 88‐Asp 89 peptide bond, observed in high‐temperature dynamics simulations, appears not to be a required step.

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C-terminal Post-translational Proteolysis of Plant Lectins and Their Recombinant Forms Expressed in Escherichia coli

Cited by 51 publications

References 33 publications

Sorting of proteins to vacuoles in plant cells

Sorting of proteins to vacuoles in plant cells

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

Structural features of the combining site region of Erythrina corallodendron lectin: Role of tryptophan 135

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