Purification of testosterone‐oestradiol‐binding globulins from mammalian sera by anion‐exchange high performance liquid chromatography

Cheng, C. Yan; Bardin, C. Wayne; Nagendranath, N; Escobar, Nereo; Han, Arthur C.; Musto, Neal A.; Gunsalus, Glen L.

doi:10.1111/j.1365-2605.1985.tb00812.x

Cited by 9 publications

(2 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The trichloroacetic acid precipitability of the [ 3 H] albumin was greater than 99%. Rabbit TeBG was purified to homogeneity, as previously described (17,18) and was tritiated with [ 3 H]NaBH 4 to a specific activity of 0.1 ixCi/ng. The [ 3 H] TeBG was 94% trichloroacetic acid precipitable and avidly bound [ 14 C]testosterone during elution from an anion exchange column (16).…”

Section: Iodination Of Imp and Tritiation Of Bovine Albumin And Rabbimentioning

confidence: 99%

Influx of Testosterone-Binding Globulin (TeBG) and TeBG-Bound Sex Steroid Hormones Into Rat Testis and Prostate*

Sakiyama

Pardridge²,

Musto³

1988

The Journal of Clinical Endocrinology & Metabolism

Self Cite

View full text Add to dashboard Cite

The availability of testosterone and estradiol to Sertoli and prostate cells is dependent upon 1) the permeability properties of the blood-tubular barrier (BTB) of the testis or prostate cell membrane, and 2) sex steroid binding to plasma proteins, such as albumin or testosterone-binding globulin (TeBG). Sex steroid influx into these tissues was studied after in vivo arterial bolus injections of [3H]testosterone or [3H]estradiol in anesthetized rats. Both testosterone and estradiol were readily cleared across the BTB or prostate cell membrane in the absence of plasma proteins and in the presence of human pregnancy serum, in which testosterone or estradiol are 80-95% distributed to TeBG. The extravascular extraction of [3H]TeBG across the BTB or prostate plasma membrane [73 +/- 2% (+/- SE) and 92 +/- 9%, respectively] was significantly greater than extraction of [3H]albumin or other plasma space markers and indicative of a rapid first pass clearance of TeBG by Sertoli or prostate cells. In summary, these studies indicate that 1) testosterone and estradiol are readily cleared by Sertoli and prostate cells; 2) albumin- and TeBG-bound sex steroids represent the major circulating pool of bioavailable hormone for testis or prostate; and 3) the TeBG-sex steroid complex may be nearly completely available for influx through the BTB or prostate plasma membrane.

show abstract

Section: Iodination Of Imp and Tritiation Of Bovine Albumin And Rabbimentioning

confidence: 99%

Influx of Testosterone-Binding Globulin (TeBG) and TeBG-Bound Sex Steroid Hormones Into Rat Testis and Prostate*

Sakiyama

Pardridge²,

Musto³

1988

The Journal of Clinical Endocrinology & Metabolism

Self Cite

View full text Add to dashboard Cite

show abstract

“…Human SHBG was purified [9,17] and photolabeled with A6-[3H]testosterone [9], and the product was analysed by SDS-PAGE and fluorography [9]. About 200/zg photolabeled SHBG was equilibrated with trypsin buffer (50 mM Tris, 30 mM KCI, 2 mM MgCI2, pH 7.5, at 22°C), 20/zg N-tosyl-L-phenylalanine chloromethyl-treated trypsin was added, and digestion was performed at 37°C for 24 h. Preliminary experiments indicated that digestion was completed using an enzyme:substrate ratio of 1:10 within 16 h under these conditions.…”

Section: Photolabeling Of Shbg and Preparation Of Tryptic Fragmentsmentioning

confidence: 99%

The cDNA‐deduced primary structure of human sex hormone‐binding globulin and location of its steroid‐binding domain

et al. 1987

View full text Add to dashboard Cite

We have sequenced a cDNA for sex hormone-binding globulin (SHBG) isolated from a phage 2gtl 1 human liver cDNA library. The library was screened with a radiolabeled rat androgen-binding protein (ABP) cDNA, and the abundance of SHBG cDNAs was 1 in 750 000 plaques examined. The largest human SHBG cDNA (1194 base-pairs) contained a reading frame for 381 amino acids. This comprised 8 amino acids of a signal peptide followed by 373 residues starting with the known NH2-terminal sequence of human SHBG, and ending with a termination codon. The predicted polypeptide Mr of SHBG is 40 509, and sites of attachment of one O-linked (residue 7) and two N-linked oligosaccharide (residues 351 and 367) chains were identified. Purified SHBG was photoaffinity-labeled with zt6-[3H]testosterone and cleaved with trypsin. The labeled tryptic fragment was isolated by reverse-phase HPLC, and its NH2-terminal sequence was determined. The results suggest that a portion of the steroid-binding domain of SHBG is located between residue 296 and the 35 predominantly hydrophilic residues at the C-terminus of the protein.cDNA cloning; Amino acid sequence; Sex hormone-binding globulin; Steroid-binding domain

show abstract

Endocytosis of Androgen‐Binding Protein, Cluster in, and Transferrin in the Efferent Ducts and Epididymis of the Ram

VEERAMACHANENI,

AMANN

1991

Journal of Andrology

View full text Add to dashboard Cite

The amount of androgen‐binding protein (ABP) In luminal fluid from the central caput epididymidis of the ram, on a per sperm basis, remains the same as that in rete testis fluid (RTF) entering the ductuli efferentes, although >85% of the testicular protein is absorbed in proximal sites. To determine if ABP is spared from endocytosis in proximal sites and if proteins are differentially and selectively absorbed at specific sites in the excurrent ducts, we studied the endocytosis of ABP, clusterin, transferrin, and a 26/35‐kd dimer isolated from ovine RTF. Each protein was labeled with colloidal gold and microinjected into the lumen of a ductulus efferens and five specific sites in the ductus epididymidis; uptake was quantified by electron microscopy. Endocytosis of each protein, including ABP, was substantially greater in the ductuli efferentes than in any site in the ductus epididymidis. More ABP was endocytosed in proximal regions of the epididymis than any other protein studied. Endocytosis of the 26/35‐kd dimer, like ABP, was greater in proximal sites of the epididymis, whereas endocytosis of clusterin and transferrin was greater in distal sites. Thus, there was a differential absorption, since proteins were endocytosed in one or another specific region of the epididymis, depending on the protein. ABP was endocytosed in the ductuli efferentes and caput epididymidis in amounts similar to or greater than other major testicular proteins, and was not spared from endocytosis in the proximal excurrent ducts. Previously reported maintenance of a similar amount of ABP, on a per sperm basis, throughout the ovine caput epididymidis probably results from recycling of endocytosed ABP or from epididymal secretion of ABP.

show abstract

Purification of testosterone‐oestradiol‐binding globulins from mammalian sera by anion‐exchange high performance liquid chromatography

Cited by 9 publications

References 25 publications

Influx of Testosterone-Binding Globulin (TeBG) and TeBG-Bound Sex Steroid Hormones Into Rat Testis and Prostate*

Influx of Testosterone-Binding Globulin (TeBG) and TeBG-Bound Sex Steroid Hormones Into Rat Testis and Prostate*

The cDNA‐deduced primary structure of human sex hormone‐binding globulin and location of its steroid‐binding domain

Endocytosis of Androgen‐Binding Protein, Cluster in, and Transferrin in the Efferent Ducts and Epididymis of the Ram

Contact Info

Product

Resources

About