Unknown

Dataset Information

0

Oxidative radioiodination damage to human lactoferrin.


ABSTRACT: Oxidative iodination of human lactoferrin (Lf) as commonly performed by using the chloramine-T, the Iodogen or the lactoperoxidase method produces an unreliable tracer protein because of excessive and heterogeneous polymer formation. Before iodination a minor tetramer fraction may be demonstrable in iron-saturated Lf only. Iodination-induced polymerization of iron-poor as well as iron-saturated Lf occurs independently of the presence or absence of 10 mM-EDTA and the 125I-/Lf molar ratio used for iodination. 125I-Lf polymers are mainly covalently linked, as suggested by the lack of substantial dissociation in SDS/polyacrylamide-gel electrophoresis. Damage to the 125I-Lf monomer may be another consequence of oxidative iodination. This is demonstrated in SDS/polyacrylamide-gel electrophoresis where 50% of the radioactivity of apparently normal monomer (Mr 75,000) is displaced to a lower-Mr region (30,000-67,000) after reduction with dithiothreitol. Non-oxidative iodination by the Bolton-Hunter technique produces an antigenetically stable tracer that is not being subjected to polymerization and monomer degradation as judged by high-performance gel chromatography and SDS/polyacrylamide-gel electrophoresis with and without dithiothreitol treatment. It is concluded that oxidation in itself leads to covalent non-disulphide cross-linking between human Lf molecules and, possibly, to intramolecular peptide-bond breaking becoming unmasked under reducing conditions. In biological experiments with human 125I-Lf this problem should be carefully considered.

SUBMITTER: Rosenmund A 

PROVIDER: S-EPMC1147399 | biostudies-other | 1986 Nov

REPOSITORIES: biostudies-other

Similar Datasets

| S-EPMC8671034 | biostudies-literature
| S-EPMC5140714 | biostudies-literature
| S-EPMC4929447 | biostudies-literature
| S-EPMC6044831 | biostudies-literature
| S-EPMC4422485 | biostudies-literature
| S-EPMC5523563 | biostudies-literature
| S-EPMC4385467 | biostudies-literature
| S-EPMC8087042 | biostudies-literature
| S-EPMC2547852 | biostudies-literature
| PRJEB11644 | ENA