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Iron‐Induced conformational change in human lactoferrin: Demonstration by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and analysis of effects of iron binding to the N and C lobes of the molecule
Author(s) -
Ying Li,
He Jianglin,
Furmanski Philip
Publication year - 1994
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150150142
Subject(s) - lactoferrin , sodium dodecyl sulfate , chemistry , gel electrophoresis , polyacrylamide gel electrophoresis , chromatography , electrophoresis , sodium , biochemistry , enzyme , organic chemistry
Analysis of Fe‐saturated‐ and apo‐lactoferrin by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) without heating the samples prior to application revealed a substantial difference in mobility. The mobility shift was fully reversible on repetitive removal and readdition of Fe. Binding of a single Fe to the N‐lobe binding site was sufficient to cause the gel shift; binding of a second Fe to the C‐lobe site did not further alter mobility. Removal of Fe from the N lobe of Fe 2 lactoferrin did not restore mobility to the position of apolactoferrin. No change in mobility with Fe binding was detected in N and C lobes isolated from intact lactoferrin by controlled trypsin digestion. The data indicate that a conformational change induced by Fe binding to a single site on lactoferrin is detectable by SDS‐PAGE and that this change requires an intact molecule, possibly due to the need for interactions between the two homologous lobes of the molecule.