Chromatography of luteinizing hormone from sheep pituitary glands

Ward, Darrell N.; Ward, Nancy E.; McGregor, Robert F.; Griffin, A. Clark

doi:10.1016/0006-3002(59)90602-x

Cited by 67 publications

(9 citation statements)

References 15 publications

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“…It has been reported that LH is heterogeneous as regards the amounts of glucosamine (Ward et al 1959) and has a few biologically active components separated by isoelectric focusing (Braselton and McShan 1970). The same biochemical properties were confirmed in human LH (Reichert 1971), bovine LH (Yoshida and Ishii 1973), rat LH (Wakabayashi 1977), and avian LH (Hattori and Wakabayashi 1979).…”

Section: Discussionsupporting

confidence: 51%

Immunoreactive luteinizing hormone (ir-LH) cells in the lung and stomach of chick embryos

Shirasawa

Shiino

Shimizu

et al. 1995

Cell and Tissue Research

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Luteinizing hormone (LH) immunoreactivity was detected in the lung and stomach of chick embryos by the immunoperoxidase staining technique using specific antiserum to chicken LH. Immunoreactive LH (ir-LH) cells first appeared in the primordial cells of the epithelial layer of lung bud and foregut as well as of Rathke s pouch in the 3-day-old embryo, Hamburger and Hamilton stage 21. Ir-LH cells increased in number with advancing age of embryos in the lung, stomach, and pituitary gland. In the lung of 7-day-old embryos, stage 31, the ir-LH cells were distributed in the epithelium of primary, secondary, and tertiary bronchi, and their shapes were pseudostratified columnar, simple columnar, and simple cuboidal, depending on their sites in the intrapulmonary airway. Ir-LH cells were more numerous in the median part than in the lateral part of the lung, and the population in the epithelial layer of entobronchi of the secondary bronchi was 4 times higher than that in ectobronchi and laterobronchi of the secondary bronchi and in the primary bronchi. The immunoreactive products were found, either in the entire cell or in the apical part, facing the lumina of bronchi. In the stomach, ir-LH cells were found in the epithelial layer of gastric glands. No ir-LH cells were observed in interstitial regions, which consisted of mesenchymal cells and blood vessels, in the lung and stomach tissues. With advancing age, ir-LH cells changed their shapes to flat or squamous, coincident with the formation of parabronchi. Other pituitary hormones were not observed immunohistochemically in either the lung or stomach before hatching. Preabsorption of the antiserum against avian LH with the purified chicken LH or the extract of pituitaries from 10-day-old embryos completely destroyed the immunoreactivity to the cells in the lung and the pituitary. A single band of the immunoreaction products, whose molecular weight was around 25 K daltons, was shown by the immunostaining of nitrocellulose membrane transblotted after sodium dodecylsulphate-polyacrylamide gel electrophoresis of the purified pituitary LH, extracts of pituitaries from 10-day-old embryos, and the extracts of lungs from 7-, 10-, and 14-day-old chick embryos. These results demonstrated that ir-LH cells are present in extrapituitary tissues, and may play an important role during the development of chick embryonic lung and stomach.

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Section: Discussionsupporting

confidence: 51%

Immunoreactive luteinizing hormone (ir-LH) cells in the lung and stomach of chick embryos

Shirasawa

Shiino

Shimizu

et al. 1995

Cell and Tissue Research

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“…purifiied certain hormones with the biological activity as an index. Working on luteinizing hormone (LH), Ward et al (1959) obtained biologically active highly purified ovine LH preparations containing different amounts of glucosamine. Braselton and McShan (1970) purified equine LH and found 4 biologically active components after electrofocusing separation.…”

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confidence: 99%

Heterogeneity of rat luteinizing hormone revealed by radioimmunoassay and electrofocusing studies.

Wakabayashi

1977

Endocrinol Japon

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Radioimmunoassay of rat luteinizing hormone (LH) in the sera and the anterior pituitaries of normal and castrated males and proestrous females employing different batches of anti-LH sera gave the results suggesting heterogeneity of immunoreactive LH (IR-LH). Gel filtration analyses of anterior pituitary extract and pooled sera failed to give positive evidence for the heterogeneity. But when the extracts of normal rat anterior pituitaries were electrofocused and the fractions were assayed, the pituitary glycoprotein hormones, LH, FSH and TSH, were found to consist of several components with different isoelectric points. In electrofocusing patterns of IR-LH, distinct peaks were observed at pH 8.5 (component A), 8.8 (component B), 9.1 (component C) and 9.3-9.4 (component D), and less distinct peaks were observed at pH 7.9 (component A'), 9.6 (component E) and 9.8 (component F). There seemed to be some sexual difference in the relative amounts of these components. Orchiectomy caused marked changes in electrofocusing patterns of IR-LH, i.e., increase of components A', A, B, C and D. The blockade of LH release by estradiol benzoate and progesterone treatment in orchiectomized rats caused a marked increase of component A. At the proestrous LH surge, IR-LH components were evenly released. Radioimmunoassay of IR-LH components with multiple antisera indicated that these antisera had different affinities to these components.

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“…The oLHa was prepared and characterized as described earlier (Ward and McGregor, 1959;Ward et al, 1967). for 4 hr at 37~ (Bousfield and Ward, 1994).…”

Section: Methodsmentioning

confidence: 99%

Circular dichroic spectroscopy of Arg46-nicked ovine lutropin α and derived fragments

et al. 1996

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The alpha subunit of ovine lutropin can be nicked with the endoproteinase Arg-C to give a single cleavage of the Arg46-Ser47 peptide bond. Following reduction by sulfitolysis, the N-terminal (residues 1-46) and C-terminal (residues 47-96) fragments can be separated and then recombined and reoxidized to yield a reconstituted nicked alpha that binds to the beta subunit but exhibits only 2-3% of the receptor-binding potency of intact lutropin. We have investigated nicked alpha, the two separated fragments, and reconstituted nicked alpha by circular dichroic spectroscopy and compared the spectra with those of intact alpha and reduced, reoxidized intact alpha. Between 200 and 225 nm the spectra of the two intact preparations are similar, as are the spectra of the two nicked preparations. However, the extremum negative ellipticities of the nicked preparations are substantially less than those of the intact preparations between 210 and 220 nm, indicating a loss in secondary structure accompanying cleavage of the Arg46-Ser47 bond. The sum of the spectra of the two fragments is significantly different from that of reconstituted nicked alpha, showing that the secondary structures in the isolated fragments are quite different from that of the reconstituted nicked protein. Reduced receptor binding by lutropin preparations containing a nicked alpha subunit may be attributable in part to the loss of secondary structure, probably helicity.

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Chromatography of luteinizing hormone from sheep pituitary glands

Cited by 67 publications

References 15 publications

Immunoreactive luteinizing hormone (ir-LH) cells in the lung and stomach of chick embryos

Immunoreactive luteinizing hormone (ir-LH) cells in the lung and stomach of chick embryos

Heterogeneity of rat luteinizing hormone revealed by radioimmunoassay and electrofocusing studies.

Circular dichroic spectroscopy of Arg46-nicked ovine lutropin α and derived fragments

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