title: Different binding affinities of Pb2+ and Cu2+ to glycosylation variants of human serum transferrin interfere with the detection of carbohydrate-deficient transferrin.
abstract: Carbohydrate-deficient transferrin (CDT) is a specific biomarker of alcohol abuse, and for diagnosis of chronic alcohol, abuse is often determined using isoelectric focusing (IEF) and chromatographic techniques. To allow this method to be used for the diagnosis of alcohol abuse, inferences of various physical and chemical factors with the detection of CDT have been investigated. However, few reports have focused thus far on whether different metal ions have different binding affinities to CDT and HTf variants or further interfere in the detection of CDT. Here, in order to figure out whether and how metal ions such as Pb(2+) and Cu(2+) bind to holo-human serum transferrin (holo-HTf) and further interfere in CDT detection, the binding characteristics and the binding parameters of holo-HTf with metal ions such as Pb(2+) and Cu(2+) were investigated using UV-visible spectroscopy, Fluorescence spectroscopy, and ICP-MS. Moreover, whether the metal ions such as Pb(2+) and Cu(2+) will reduce the diagnostic accuracy of CDT in clinic was investigated using IEF. The present study demonstrates that Pb(2+) and Cu(2+) have different binding affinities to holo-HTf variants and produce different changes in the relative amounts of each glycosylation isoforms of HTf. Accordingly, the glycosylation chains of HTf will affect the binding affinities of glycosylation isoforms with Pb(2+) and Cu(2+), causing further interferences in CDT detection.
authors.affiliation: Key Laboratory of MOE for Cell Biology and Tumor Cell Engineering, School of Life Sciences, Xiamen University, Xiamen, 361005, China.
authors.forename: Lian-Zhong
authors.initials: LZ
authors.lastname: Luo
authors.forename: Hong-Wei
authors.initials: HW
authors.lastname: Jin
authors.forename: Lin
authors.initials: L
authors.lastname: Huang
authors.forename: He-Qing
authors.initials: HQ
authors.lastname: Huang
mesh_terms: D015415:Biomarkers; D003300:Copper / Q000097:blood*; D005189:False Positive Reactions; D006031:Glycosylation; D006801:Humans; D007525:Isoelectric Focusing; D007854:Lead / Q000097:blood*; D013058:Mass Spectrometry; D011485:Protein Binding; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D014168:Transferrin / Q000031:analogs & derivatives* / Q000032:analysis / Q000302:isolation & purification
chemical_list: D015415:Biomarkers; D014168:Transferrin; C056759:carbohydrate-deficient transferrin; D007854:Lead; D003300:Copper