Comparison of two methods for radioiodination on the oxidizability properties of low density lipoprotein.
Radiolabeling of low density lipoprotein (LDL) apoB100 with 125I, an oxidative process, is commonly used in lipoprotein investigation. Since 1) LDL is unstable and oxidation-prone, 2) the modification of apoB100 by oxidation increases the negative charge of particles and leads to the uptake of modified LDL through the scavenger receptor pathway, and 3) oxidized LDL is cytotoxic, it is relevant to investigate whether the oxidative stability of LDL is influenced by its labeling with 125I. The aim of this study was to investigate and compare lipid and protein oxidation markers in human LDL after labeling with 125I by two widely adopted methods that use ICl or the chloramide 1,3,4,6-tetrachloro-3alpha,6alpha-diphenylglycoluril as the oxidizing agent. Native LDL served as a common control and sham-iodinated LDL as a handling control for each procedure. The resistance against copper-induced oxidation of 125I-LDL labeled with ICl was similar to that of controls with regard to the lag time and maximal amount of conjugated diene formed, as there were levels of initial conjugated diene, alpha-tocopherol, and tryptophan. However, radioiodination with the chloramide accelerated the onset of the rapid phase of LDL oxidation due to a drastic depletion of alpha-tocopherol and increased conjugated diene content. Measurements of copper-induced LDL oxidizability showed enhanced indices of lipid oxidation. The lag time and the time to maximal diene production were 65% and 30% shorter than controls. This was accompanied by a 50% reduced tryptophan fluorescence. The anionic surface charge of the LDL particle increased moderately with both labeling procedures. The results indicate that labeling of LDL with 125I may oxidize lipids and apoB100 to a variable extent, depending on the nature of the iodinating agent. This is why assessment of the oxidizability properties of 125I-labeled LDL is recommended for reliable biological studies.
Romero
J R
JR
Department of Physiology, University of the Basque Country Medical School, Bilbao, Spain.
Martínez
R
R
Fresnedo
O
O
Ochoa
B
B
D000328:Adult; D000975:Antioxidants / Q000378:metabolism; D002712:Chlorides / Q000378:metabolism; D003300:Copper / Q000378:metabolism; D006801:Humans; D007202:Indicators and Reagents / Q000378:metabolism; D007454:Iodides / Q000378:metabolism; D007457:Iodine Radioisotopes / Q000378:metabolism*; D007553:Isotope Labeling / Q000379:methods*; D008077:Lipoproteins, LDL / Q000378:metabolism* / Q000528:radiation effects; D010084:Oxidation-Reduction; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism; D014364:Tryptophan / Q000378:metabolism; D014508:Urea / Q000031:analogs & derivatives* / Q000378:metabolism; D024502:alpha-Tocopherol / Q000378:metabolism
D000975:Antioxidants; D002712:Chlorides; D007202:Indicators and Reagents; D007454:Iodides; D007457:Iodine Radioisotopes; D008077:Lipoproteins, LDL; D017392:Thiobarbituric Acid Reactive Substances; C099252:oxidized low density lipoprotein; C009813:iodine monochloride; C012385:1,3,4,6-tetrachloro-3 alpha,6 alpha-diphenylglycoluril; D003300:Copper; D014364:Tryptophan; D014508:Urea; D024502:alpha-Tocopherol