Radioiodination of chicken luteinizing hormone without affecting receptor binding potency.
By improving the currently used lactoperoxidase method, we were able to obtain radioiodinated chicken luteinizing hormone (LH) that shows high specific binding and low nonspecific binding to a crude plasma membrane fraction of testicular cells of the domestic fowl and the Japanese quail, and to the ovarian granulosa cells of the Japanese quail. The change we made from the original method consisted of 1) using chicken LH for radioiodination that was not only highly purified but also retained a high receptor binding potency; 2) controlling the level of incorporation of radioiodine into chicken LH molecules by employing a short reaction time and low temperature; and 3) fractionating radioiodinated chicken LH further by gel filtration using high-performance liquid chromatography. Specific radioactivity of the final 125I-labeled chicken LH preparation was 14 microCi/micrograms. When specific binding was 12-16%, nonspecific binding was as low as 2-4% in the gonadal receptors. 125I-Labeled chicken LH was displaced by chicken LH and ovine LH but not by chicken follicle-stimulating hormone. The equilibrium association constant of quail testicular receptor was 3.6 x 10(9) M-1. We concluded that chicken LH radioiodinated by the present method is useful for studies of avian LH receptors.
Kikuchi
M
M
Department of Biology, School of Education, Waseda University, Tokyo, Japan.
Ishii
S
S
D000818:Animals; D002645:Chickens / Q000502:physiology*; D003370:Coturnix / Q000502:physiology*; D005260:Female; D007457:Iodine Radioisotopes; D007986:Luteinizing Hormone / Q000032:analysis / Q000378:metabolism*; D008297:Male; D011784:Quail / Q000502:physiology*; D011863:Radioimmunoassay; D011869:Radioligand Assay; D011974:Receptors, LH / Q000378:metabolism*
D007457:Iodine Radioisotopes; D011974:Receptors, LH; D007986:Luteinizing Hormone