Unfolding of iron and copper complexes of human lactoferrin and transferrin

Harrington, John P.; Stuart, Jan; Jones, A.

doi:10.1016/0020-711x(87)90184-4

Cited by 15 publications

(5 citation statements)

References 39 publications

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“…This is because only one highly cooperative transition was observed for the SUPREX curves obtained for apotransferrin (Figure A−C). This is consistent with the results of earlier studies in which one cooperative unfolding transition was observed for apotransferrin in a conventional GdmCl denaturation experiment. , The relatively large m -values determined for the fragments in our SUPREX-protease digestion experiments on apotransferrin are also consistent with the increased cooperativity that would be expected if the two lobes of apotransferrin simultaneously unfolded in the GdmCl-induced equilibrium unfolding reaction.…”

Section: Discussionsupporting

confidence: 92%

H/D Exchange and Mass Spectrometry-Based Method for Biophysical Analysis of Multidomain Proteins at the Domain Level

2007

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A protocol was developed to characterize the domain-specific thermodynamic stabilities of multidomain proteins using SUPREX (Stability of Unpurified Proteins from Rates of H/D Exchange). The protocol incorporates a protease digestion step into the conventional SUPREX protocol and enables folding free energy (DeltaGf) and cooperativity (m-value) measurements to be made on the individual domains of multidomain proteins in their native context (i.e., in the intact protein). Three multidomain protein systems (calmodulin, a Fyn construct, and transferrin) were used to validate the SUPREX-protease digestion protocol. The DeltaGf and m-value of each domain in the intact test proteins were measured in the absence and presence of ligands using the new protocol. Domain-specific thermodynamic parameters were obtained on each system; and the measured parameters were consistent with known biophysical properties of the test proteins. The known stabilization of the N-terminal domain of CaM in the context of the intact protein and the known binding affinity of a proline-rich peptide to the SH3 domain in the Fyn construct were successfully quantified using the new protocol. Qualitative information about the relative calcium binding affinities of the N- and C-terminal domains of CaM and about the relative iron binding affinities of the N- and C-terminal domains of transferrin was also obtained using the new protocol.

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Section: Discussionsupporting

confidence: 92%

H/D Exchange and Mass Spectrometry-Based Method for Biophysical Analysis of Multidomain Proteins at the Domain Level

2007

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“…With the apotransferrin, the thermodynamic equilibrium is reached very rapidly, whereas with hololactoferrin the ®nal equilibrated state is attained a few thousand seconds later after iron loss (Figure 1). This implies, as already reported, that the iron load protects the transferrin from denaturation (Harrington et al, 1987;Harrington, 1992). Indeed, hololactoferrin keeps its iron-binding capacities even after mild tryptic digestion (Legrand et al, 1984).…”

Section: Discussionsupporting

confidence: 76%

“…This re¯ects a common change in conformation, which may be a partial unfolding of lactoferrin. Folding and unfolding of transferrins are not well known and, to our knowledge, the kinetics of unfolding of these proteins have not been investigated (Morgan & Peters, 1985;Harrington et al, 1987;Harrington, 1992;Lodish & Kong, 1991;Kilar & Hjerten, 1993;Ou et al, 1993;Wada et al, 1997). This change in conformation affects both apo-and holotransferrin.…”

Section: Discussionmentioning

confidence: 99%

Transferrins: iron release from lactoferrin

Abdallah

Chahine

2000

Journal of Molecular Biology

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“…The iron-binding capacity of lactoferrin is dependent on the presence of (small amounts) of (bi)carbonate. The binding site appears to be optimized for the binding of ferric iron and (bi)carbonate with respect to size, charge and stereochemistry, as evidenced from a number of structural studies with different anions and cations, or using mutant recombinant lactoferrins (Harrington et al 1987;Baker et al 1994;Brodie et al 1994;Faber et al 1997). Oxalate can replace (bi)carbonate with regard to iron binding, but citrate can not.…”

Section: Iron Bindingmentioning

confidence: 99%

Occurrence, structure, biochemical properties and technological characteristics of lactoferrin

Steijns¹,

Hooijdonk²

2000

Br J Nutr

264

177

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The structure of the iron-binding glycoprotein lactoferrin, present in milk and other exocrine secretions, has been elucidated in great detail, both the three-dimensional protein structure and the attached N-glycans. Structure±function relationships are being established. From these studies a function for lactoferrin in host defence and modulation of iron metabolism emerges. This paper describes in some detail how iron and other cations may be bound by lactoferrins from human or bovine sources and elucidates parts of the molecule that are critical for interactions with cells and biomolecules. Furthermore, the technological aspects, more specifically the heat-sensitivity, of bovine lactoferrin in different matrices are described.

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Unfolding of iron and copper complexes of human lactoferrin and transferrin

Cited by 15 publications

References 39 publications

H/D Exchange and Mass Spectrometry-Based Method for Biophysical Analysis of Multidomain Proteins at the Domain Level

H/D Exchange and Mass Spectrometry-Based Method for Biophysical Analysis of Multidomain Proteins at the Domain Level

Transferrins: iron release from lactoferrin

Occurrence, structure, biochemical properties and technological characteristics of lactoferrin

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