ISOLATION OF HIGHLY PURIFIED SEX HORMONE BINDING GLOBULIN (SHBG): EVIDENCE FOR MICROHETEROGENEITY

1979 ◽  
Vol 90 (4) ◽  
pp. 737-742 ◽  
Author(s):  
W. Mischke ◽  
H. C. Weise ◽  
D. Graesslin ◽  
R. Rusch ◽  
J. Tamm
Keyword(s):  
Human Serum ◽  
Isoelectric Focusing ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Double Diffusion ◽  
Polyacrylamide Gel ◽  
Sex Hormone ◽  
Sex Hormone Binding Globulin ◽  
Hormone Binding ◽  
Gel Isoelectric Focusing

ABSTRACT Highly purified sex hormone binding globulin (SHBG) was isolated in milligram amounts from a human serum fraction (Cohn IV-4). The final preparation was homogeneous by the criteria of polyacrylamide-gel electrophoresis. Immunological evidence for purity could be given by double diffusion according to Ouchterlony. However, following gel isoelectric focusing highly purified SHBG displayed four different bands, as could be demonstrated by staining as well as by a photoscan of the [3H]5α-dihydrotestosterone-SHBG complex. After incubation with neuraminidase the microheterogeneity of SHBG disappeared and the asialo-SHBG showed only one band.

scholarly journals The rapid purification and partial characterization of human serum angiotensinogen

1987 ◽  
Vol 243 (1) ◽  
pp. 121-126 ◽  
Author(s):  
C J Campbell ◽  
P A Charlton ◽  
C J Grinham ◽  
C J Mooney ◽  
J E Pendlebury
Keyword(s):  
Sialic Acid ◽  
Human Serum ◽  
Isoelectric Focusing ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Multiple Forms ◽  
Rapid Purification ◽  
Blue Sepharose

Human angiotensinogen has been purified 390-fold from serum by a rapid high-yielding procedure that involved chromatography on Blue Sepharose, phenyl-Sepharose, hydroxyapatite and immobilized 5-hydroxytryptamine (5-HT). Angiotensinogen was specifically bound to immobilized 5-HT, which effected a partial resolution into multiple forms, which were also evident when analysed by SDS/polyacrylamide-gel electrophoresis (Mr 59,400, 60,600, 62,600 and 63,800). This heterogeneity was confirmed by resolution into six main bands on isoelectric focusing, ranging from pI 4.40 to 4.82. N-terminal analysis, digestion with human renal renin and deglycosylation studies implied that the preparation comprised several forms of angiotensinogen, varying in their degree of glycosylation. The presence of sialic acid was shown to be a major factor in determining the heterogeneity.

Genetic polymorphism of the human sex hormone-binding globulin: Evidence of an isoelectric focusing variant with normal androgen-binding affinities

1990 ◽  
Vol 36 (6) ◽  
pp. 541-548 ◽  
Author(s):  
Fernando Larrea ◽  
Rosa Maria Oliart ◽  
Julio Granados ◽  
Osvaldo Mutchinick ◽  
Vicente Diaz-Sanchez ◽  
...  
Keyword(s):  
Genetic Polymorphism ◽  
Isoelectric Focusing ◽  
Sex Hormone ◽  
Sex Hormone Binding Globulin ◽  
Binding Affinities ◽  
Hormone Binding ◽  
Androgen Binding

Microheterogeneity of sex hormone binding globulin isolated from a human serum fraction

1977 ◽  
Vol 8 (12) ◽  
pp. xviii
Author(s):  
W. Mischke ◽  
H.C. Weise ◽  
D. Graesslin ◽  
J. Tamm
Keyword(s):  
Human Serum ◽  
Sex Hormone ◽  
Sex Hormone Binding Globulin ◽  
Hormone Binding

Binding of Testosterone and Oestradiol to Sex Hormone Binding Globulin, Human Serum Albumin and other Plasma Proteins: Evidence for Non-Specific Binding of Oestradiol to Sex Hormone Binding Globulin

1983 ◽  
Vol 64 (3) ◽  
pp. 307-314 ◽  
Author(s):  
M. Jawed Iqbal ◽  
Maureen Dalton ◽  
Robert S. Sawers
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Binding Site ◽  
Plasma Proteins ◽  
Specific Binding ◽  
Sex Hormone ◽  
Sex Hormone Binding Globulin ◽  
Normal Female ◽  
Hormone Binding

1. The percentage binding of testosterone (T) and oestradiol (E2) to sex hormone binding globulin (SHBG) and human serum albumin (HSA) was determined over a range of SHBG concentrations of 16–250 nmol of dihydrotestosterone (DHT) bound/l. It was found that the binding of both T and E2 to HSA was a function of their binding to SHBG and bore an inverse relationship to it. After removal of both SHBG and HSA from plasma by affinity chromatography a ‘residual’ binding of about 11% for T and 12% for E2 was still apparent. in addition to the specific high-affinity, low capacity binding of E2 to SHBG, non-specific low-affinity binding of 7–12% was demonstrated after selective denaturation of the specific binding site of the latter. 2. Competition studies indicated that although at the relatively higher levels of SHBG found in the normal female the physiological concentrations of E2, T and DHT need not be taken into account in estimating the unbound fractions of steroids, at the relatively lower levels of SHBG found in normal men and hirsute women, the physiological concentrations of T and DHT are effective in causing statistically significant displacement of E2 from the common, specific binding site on SHBG. 3. A simple computerized technique is described for the determination of fractions of E2 and T respectively, that are unbound to SHBG, unbound to SHBG and HSA, and unbound to all plasma proteins, when the total plasma levels of E2, T, DHT and SHBG are known.

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