scholarly journals The Effect of Oxidation and pH on the Molecular Weight of Human Growth Hormone, Estimated by Gel Filtration

1966 ◽  
Vol 101 (3) ◽  
pp. 43C-44C ◽  
Author(s):  
P. Fønss-Bech
Keyword(s):  
Molecular Weight ◽  
Growth Hormone ◽  
Human Growth Hormone ◽  
Gel Filtration ◽  
Human Growth

Effects of glycosylation inhibitors on human growth hormone receptor in cultured human lymphocytes

1988 ◽  
Vol 119 (4) ◽  
pp. 517-524 ◽  
Author(s):  
Kumiko Asakawa ◽  
Jose A. Hedo ◽  
Phillip Gorden ◽  
Kazuo Shizume
Keyword(s):  
Molecular Weight ◽  
Growth Hormone ◽  
Human Growth Hormone ◽  
Growth Hormone Receptor ◽  
Human Lymphocytes ◽  
Human Growth ◽  
Control Value ◽  
Receptor Complexes ◽  
Oligosaccharide Processing ◽  
Receptor Number

Abstract. IM-9 cultured human lymphocytes were treated with N-linked glycosylation inhibitors, N-linked oligosaccharide processing inhibitors, or neuraminidase to study the effect of glycosylation modification on human growth hormone binding and molecular weight of surface hGH receptor. One mg/l tunicamycin and 20 mmol/l glucosamine decreased 125I-hGH binding to the cells to 46.3 ± 2.4% (mean ± sem) and 21.9 ± 0.2% of the controls, respectively. The hGH binding was 33.0 ± 18.4% of the control value in the cells treated with monensin. The inhibition of binding was due to a decrease in the hGH receptor number without any affinity changes in these cells. Neither 1 mg/l swainsonine nor 100 mg/l castanospermine had any effect on the hGH binding. On the other hand, 125I-hGH binding to neuraminidase-treated cells was significantly enhanced with accompanying affinity changes. When 125I-hGH was cross-linked to IM-9 cells, there were no differences in the molecular weight of hGH receptor complexes (140K) between untreated cells and cells treated with tunicamycin, glucosamine, monensin, or castanospermine. However, the 128K hGH-receptor complex appeared in swainsonine-treated cells; this complex was sensitive to endoglycosidase H. These data show that the altered carbohydrate moiety changed hGH binding and the size of surface hGH receptor and suggest that glycosylation of receptor is important for the binding of hGH and for its physiological action.

Conversion of Radiolabeled Human Growth Hormone into Higher Molecular Weight Moieties in Human Plasmain Vivoandin Vitro

Endocrinology ◽  
1977 ◽  
Vol 101 (2) ◽  
pp. 350-359 ◽  
Author(s):  
INESE Z. BEITINS ◽  
MARIO C. RATTAZZI ◽  
MARGARET H. MACGILLIVRAY
Keyword(s):  
Molecular Weight ◽  
Growth Hormone ◽  
Human Growth Hormone ◽  
Human Growth

IMMUNOREACTIVE GROWTH HORMONE IN HUMAN URINE

1972 ◽  
Vol 71 (4) ◽  
pp. 665-676 ◽  
Author(s):  
Kristian F. Hanssen
Keyword(s):  
Molecular Weight ◽  
Growth Hormone ◽  
Linear Relationship ◽  
Human Urine ◽  
Gel Filtration ◽  
Human Growth ◽  
Normal Subjects ◽  
Normal Human ◽  
Radio Immunoassay

ABSTRACT By using a double antibody radio-immunoassay (pre-precipitation technique) for the determination of immunoreactive human growth hormone (IRHGH) in normal human urine concentrated by dialysis and lyophilization, a factor was revealed that displaces 125I-HGH from HGH antibodies. This displacement was neither due to salts nor to glucose; it is suggested that it is due to IRHGH in the urine. A linear relationship between dilution of urine and the measured IRHGH concentration was obtained. Recovery of exogenous HGH was between 70–105%. The recovery of IRHGH from different volumes of urine following dialysis and lyophilization was between 97–110%. Plasma IRHGH and urinary IRHGH was measured simultaneously after HGH injection in a normal subject. A correlation was shown between plasma IRHGH and urinary IRHGH. In 9 normal subjects, the urinary IRHGH ranged from 28–53 ng/24 h. The excretion of urinary IRHGH was increased in acromegaly and was diminished in some, but not in all patients with adult hypopituitarism. The urinary IRHGH was further studied by gel filtration. It was recovered in one peak corresponding to a molecular weight of approximately 20 000 – 30 000. However, in the present work it was not clarified whether the urinary IRHGH represents pituitary HGH excreted in the urine or a metabolite of high molecular weight with retained immunological properties.

Radioimmunoassay of Human Growth Hormone

1968 ◽  
Vol 14 (2) ◽  
pp. 145-155 ◽  
Author(s):  
Mechthilde Knoller ◽  
M U Tsao ◽  
George H Lowrey
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Gel Filtration ◽  
Human Growth ◽  
Reproducible Method

Abstract Detailed methods for: (1) 131I iodination of human growth hormone (HGH); (2) purification of labeled HGH (HGH-131I; and (3) radioimmunoassay of HGH are given. Bio-Gel filtration is introduced as a rapid and reproducible method for purifying HGH-131I which has been obtained by a modification of the method of Greenwood et al. (1). Highly purified HGH-131I with a bindability of 96% or more is usually achieved.

scholarly journals Analysis of binding properties between 20 kDa human growth hormone (hGH) and hGH receptor (hGHR): the binding affinity for hGHR extracellular domain and mode of receptor dimerization

1999 ◽  
Vol 23 (3) ◽  
pp. 347-353 ◽  
Author(s):  
H Uchida ◽  
S Banba ◽  
M Wada ◽  
K Matsumoto ◽  
M Ikeda ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Cell Proliferation ◽  
Human Growth Hormone ◽  
Gel Filtration ◽  
Human Growth ◽  
Extracellular Domain ◽  
Experimental Result ◽  
Receptor Dimerization ◽  
Sequential Binding ◽  
D Values

It has recently been shown that 20 kDa human growth hormone (hGH) forms the 1:2 hGH:hGH receptor (hGHR) complex and expresses full agonistic activity, although it hardly forms the 1:1 GH:GHR complex as compared with 22 kDa hGH. To clarify this mechanism, we analyzed the mode of receptor dimerization of 20 kDa hGH using the intact form and mutants. Complex formation analysis between hGHR extracellular domain (hGHBP) and either site1 mutant (K157A) or site2 mutant (G105R) by gel-filtration showed that the site1 mutant apparently formed no 1:1 complex and that the site2 mutant formed only the 1:1 complex. Cell proliferation analysis revealed that the activity curve (vs ligand concentration) of 20 kDa hGH showed a bell-shaped pattern. This indicates that the receptor dimerization of 20 kDa hGH proceeds in a sequential manner. Based on this sequential binding we have produced a mathematical model for receptor dimerization as a function of [hGH], [hGHBP], K(d) values for the first hGHBP binding (K(d1)) and the second hGHBP binding (K(d2)). The result of 20 kDa hGH binding to (S201C) hGHBP immobilized on biosensor tip showed that the K(d1) value was 1. 6x10(-8) M. Adopting this value as a constant in the function described above, we have obtained calculative hGHR dimerization curves vs hGH concentration. Since the K(d2) value could not be experimentally determined, the curves were simulatively obtained with varied K(d2) values. The simulated curve pattern coincided with the experimental result of the cell proliferation in Ba/F3-hGHR when the value 2.5x10(-10) M was adopted as K(d2). In conclusion, although the affinity of 20 kDa hGH for the first hGHR binding is reduced to one-tenth, that for the second binding is increased ten-fold in comparison with those of 22 kDa hGH, indicating that 20 kDa hGH can be an effective hGH isoform in the presence of hGHBP.

Preparation of human growth hormone by gel filtration

1963 ◽  
Vol 74 ◽  
pp. 525-531 ◽  
Author(s):  
P. Roos ◽  
H.R. Fevold ◽  
C.A. Gemzell
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Gel Filtration ◽  
Human Growth

Optimization of Radioimmunoassay for Human Growth Hormone by the Charcoal-Dextran Technique

1970 ◽  
Vol 16 (10) ◽  
pp. 845-848 ◽  
Author(s):  
Joseph C Meek ◽  
Mary M Stoskopf ◽  
Robert E Bolinger
Keyword(s):  
Molecular Weight ◽  
Growth Hormone ◽  
Guinea Pig ◽  
Human Growth Hormone ◽  
Serum Proteins ◽  
Human Growth ◽  
Molecular Fragments ◽  
Antibody Technique ◽  
Double Antibody

Abstract The effect of serum on the radioimmunoassay for human growth hormone was tested by use of the double-antibody technique and the charcoal-coated dextran technique. With the double-antibody technique, serum effects a falsely high reading for unknowns, while with the charcoal-dextran technique it causes falsely low values to be obtained. Part of the anomaly with the charcoal-dextran technique is the result of adsorption of small molecular fragments of radiolabeled hormone onto serum proteins of higher molecular weight. These fragments can be eliminated by frequent repurification of the labeled hormone. Another part is not related to purity of the label but is corrected by subtracting values for serum blanks that contain no antibody. The use of antiserum to human growth hormone prepared in the goat is more sensitive than that prepared in the guinea pig.

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