Amit T, Barkey R J, Youdim M B, Hochberg Z
Rappaport Family Institute for Research in the Medical Sciences, Faculty of Medicine, Technion, Haifa, Israel.
J Clin Endocrinol Metab. 1990 Aug;71(2):474-80. doi: 10.1210/jcem-71-2-474.
The recent discovery of a GH-binding protein (GH-BP) in human serum has important implications for the distribution, metabolism, and physiological activity of human GH (hGH). In this report we describe an accurate, simple, and rapid assay for measuring GH-BP in human serum, based on the dextran-coated charcoal technique for separating bound from free hGH. The present method reduces the degree of dissociation of the GH-BP complex during the separation procedure, thus increasing accuracy, allows routine use of small working volumes of sera, and can be practically used for running large scale assays. Binding of [125I]hGH to human serum GH-BP, identified and characterized by this procedure, was time, temperature, and dose dependent; the binding was highly specific for hGH and with a high affinity (Ka, 1.08 +/- 0.19 x 10(9) L/mol). Preliminary results, expressed as percent specifically bound [125I]hGH corrected for endogenous hGH, indicate that the mean GH-BP activity in 50 microL sera from normal adult men and women was 11.32 +/- 0.45%. Significantly lower values were obtained in sera of infants, 3 days after birth (1.65 +/- 0.15%), and in short-statured children with normal GH secretion (7.38 +/- 0.22%). In patients with liver cirrhosis GH-BP levels decreased to 5.93 +/- 1.14%. The availability of a simple and convenient procedure for large scale determination of GH-BP coupled with the suggestion that this protein can reflect the GH receptor provides us with an indirect means for assessing changes in GH receptor activity in various physiological and pathological conditions.
最近在人血清中发现生长激素结合蛋白(GH - BP),这对人生长激素(hGH)的分布、代谢及生理活性具有重要意义。在本报告中,我们描述了一种基于葡聚糖包被活性炭技术分离结合态与游离态hGH的准确、简便且快速的检测人血清中GH - BP的方法。本方法在分离过程中降低了GH - BP复合物的解离程度,从而提高了准确性,允许常规使用少量血清工作体积,并且可实际用于大规模检测。通过该方法鉴定和表征的[125I]hGH与人血清GH - BP的结合具有时间、温度和剂量依赖性;该结合对hGH具有高度特异性且亲和力高(Ka,1.08±0.19×10⁹L/mol)。以校正内源性hGH后特异性结合的[125I]hGH百分比表示的初步结果表明,正常成年男性和女性50μL血清中的平均GH - BP活性为11.32±0.45%。出生3天的婴儿血清(1.65±0.15%)以及生长激素分泌正常的身材矮小儿童血清(7.38±0.22%)中的值明显较低。肝硬化患者的GH - BP水平降至5.93±1.14%。一种用于大规模测定GH - BP的简单便捷方法的可用性,以及该蛋白可反映生长激素受体的提示,为我们提供了一种间接手段来评估各种生理和病理条件下生长激素受体活性的变化。