Abstract
The aim of the study was to illustrate the radio-receptor assay of beetal recombinant caprine growth hormone (rcGH). Tracer ( 125 I-rcGH) was prepared by iodinating beetal rcGH with iodine-125 and its biological activity was analyzed by rabbit anti-rcGH antibodies. Liver microsomal membranes of the Bovidae species (caprine, ovine and bovine) were prepared to study the binding properties of beetal rcGH. The tracer binding was dependent on receptor protein concentration, tracer counts, temperature, time of incubation and assay pH. The maximum percentage specific binding of the tracer to the microsomal membrane was 45% in 32 h at 4°C. In cross-reactive study, human GH competed and displaced the 125 I-rcGH by binding effectively to the cGH receptor. Scatchard analysis of the rcGH binding suggested a single class of binding site to the caprine microsomal membrane with an affinity of 337.1 ± 82.94 × 10 9 M -1 and capacity of 57.61 ± 4.23 fmol mg -1 while rcGH binding affinity and capacity to the ovine and bovine receptor protein showed 365.4 ± 66.82 × 10 9 M -1 , 57.69 ± 3.23 (fmol mg -1 ) and 392.6 ± 56.25 × 10 9 M -1 , 56.8 ± 2.56 (fmol mg -1 ) respectively. This study provides data for beetal rcGH interaction with microsomal membrane that shall be beneficial to study hormone receptor interactions of other Bovidae species. Keywords: Beetal recombinant caprine growth hormone, Bovidae, iodine-125, microsomal membrane, receptor protein African Journal of Biotechnology , Vol 13(30) 3081-3091
Highlights
Growth hormone (GH) a 22 kDa polypeptide is produced by somatotroph cells of the anterior pituitary gland and its mode of action is widespread (Edmondson et al, 2003).It is accepted that receptors for the GH are widely occurring on the cell membrane and crude membrane preparations of various tissues (Posner et al, 1974)
This study provides data for beetal recombinant caprine growth hormone (rcGH)
Recombinant beetal cGH was produced in the laboratory (School of Biological Sciences, University of the Punjab, Lahore, Pakistan) and human GH used for the cross reactive study was a gift from NETRIA, UK
Summary
Growth hormone (GH) a 22 kDa polypeptide is produced by somatotroph cells of the anterior pituitary gland and its mode of action is widespread (Edmondson et al, 2003). It is accepted that receptors for the GH are widely occurring on the cell membrane and crude membrane preparations of various tissues (Posner et al, 1974). The most specific binding site for the GH is the liver, where it acts and produces insulin-like growth. Factors (IGF-I and II), which targets other tissues of the body (adipose tissues, bones, muscles) leading to cell proliferation and increases the protein, lipid and carbohydrate metabolism (Leung and Ho, 2001). The initial step in studying the mechanism of signal transduction of a hormone (Piwien-Pilipuk et al, 2002)
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