Isolation and Partial Characterization of LH, FSH and TSH from Canine Pituitary Gland.

Chiba, Koji; Kobayashi, Hill Hiroki; Wakabayashi, Keiji

doi:10.1507/endocrj.44.205

Cited by 4 publications

(1 citation statement)

References 36 publications

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“…[17][18][19][20] To reduce the number of steps, pseudo affinity or immunoaffinity chromatographies have also been incorporated. 17,[20][21][22][23][24] In several cases of pseudo affinity chromatography, the interaction of thyrotropin family of hormones with triazine dyes was exploited. 18,20,22,23,25 Out of several triazine dyes available, cibacron blue F3GA has been used as pseudo affinity ligand by many workers for purification.…”

Section: Introductionmentioning

confidence: 99%

Interaction Between Cibacron Blue F3GA and Luteinizing Hormone: A Chromatographic Investigation

Chaudhary¹

2012

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Luteinizing hormone which was bound to Cibacron Blue F3GA could not be eluted with 10mM NAD. Bound LH could be eluted partially (up to 5%) with 80% ethylene glycol and rest with 50mM phosphate buffer pH 7.5 containing 1M NaCl. Each of the bound fractions could also be sub-fractionated with differential elution with a gradient of ethylene glycol or NaCl. This indicated that pituitary LH was a mixture of two different sub-populations of LH, one which interacts with CB predominantly via hydrophobic interactions and the other via electrostatic interaction. In the case of subunits of LH that occur in free state in pituitaries, approximately half of the bound subunits interacted with CB column predominantly via hydrophobic interactions whereas the other half interacted via electrostatic force. It is concluded that differences in glycan content, composition and structure could be the cause for differential binding of buLH and its free subunits to the CBA column. 243 | P a g e luteinizing hormone. Results indicate that under appropriate conditions of elution, heterodimeric LH can be fractionated into different isoforms. It has also been shown here that the dye exhibits different binding preferences for the two free subunits (alpha and beta) of buLH.

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Section: Introductionmentioning

confidence: 99%

Interaction Between Cibacron Blue F3GA and Luteinizing Hormone: A Chromatographic Investigation

Chaudhary¹

2012

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Caprine (Capra hircus) Luteinizing Hormone: Purification and Chromatographic Investigation of its Different Isoforms

Chaudhary

Muralidhar

2007

Preparative Biochemistry and Biotechnology

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Luteinizing Hormone (LH) from goat pituitary glands has been purified and characterized with respect to its size and subunit nature. The purification at each step was monitored by protein estimation, SDS-PAGE, and direct binding ELISA. The final product was found to be over 90 fold purified as compared to the starting pituitary extract, and the yield of the final purified LH was found to be 65.3 mg/kilogram of wet pituitary glands. Fractionation of the cLH into different charge isoforms by SP-Sephadex ion exchanger has been observed. Chromatography on immobilized Con A lectin resulted in fractionation of the purified cLH into unbound (2%), loosely bound (85%), and firmly bound (13%) fractions indicating oligosaccharide heterogeneity. The purified hormone was capable of stimulating weight increase in the seminal vesicles in immature male rats, with a biopotency equivalent to the 2200 I.U. of hCG per mg of purified cLH. The FSH content of the purified cLH was found to be less than 0.0165% as indicated by in vivo Steelman-Pohley assay.

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Characterization, Localization, and Stage-Dependent Gene Expression of Gonadotropin Receptors in Chub Mackerel (Scomber japonicus) Ovarian Follicles

Nyuji

Kitano

Shimizu

et al. 2013

Biology of Reproduction

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The pituitary gonadotropins (GtHs), follicle-stimulating hormone (FSH) and luteinizing hormone (LH), are key regulators of gametogenesis in teleosts. However, little is known about the physiological mechanisms by which GtHs regulate asynchronous oocyte development in multiple-spawning marine fishes. We cloned cDNAs encoding GtH receptors (FSHR and LHR) from chub mackerel (Scomber japonicus). FSH and LH were purified by anion-exchange chromatography, gel filtration, and concanavalinA-agarose. When expressed in mammalian cells, FSHR and LHR responded strongly to their own ligands. By separating LH into two subunits by the use of reverse-phase chromatography, we found that the beta-subunit is responsible for signal transduction and the alpha-subunit may be important for holding hormone-receptor complex. In situ hybridization showed that only fshr was expressed in prefollicle and granulosa cells in oocytes at the perinucleolus and cortical alveolus stages, suggesting that FSH is involved in the primary and early secondary growth of oocytes. In ovarian follicles during vitellogenesis, both fshr and lhr were expressed in granulosa and thecal cells, and lhr was strongly expressed during germinal vesicle migration (GVM). Real-time PCR analysis of stage-dependent fshr and lhr expression showed that fshr expression was high in ovarian follicles throughout vitellogenesis and decreased during GVM, whereas lhr expression was low in early vitellogenesis, but increased markedly in the late phase of vitellogenesis, remaining high during GVM. These findings suggest that switching of the expression of FSHR to LHR controls the effects of FSH and/or LH on vitellogenesis and final oocyte maturation via steroid production in granulosa and thecal cells.

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Isolation and Partial Characterization of LH, FSH and TSH from Canine Pituitary Gland.

Cited by 4 publications

References 36 publications

Interaction Between Cibacron Blue F3GA and Luteinizing Hormone: A Chromatographic Investigation

Interaction Between Cibacron Blue F3GA and Luteinizing Hormone: A Chromatographic Investigation

Caprine (Capra hircus) Luteinizing Hormone: Purification and Chromatographic Investigation of its Different Isoforms

Characterization, Localization, and Stage-Dependent Gene Expression of Gonadotropin Receptors in Chub Mackerel (Scomber japonicus) Ovarian Follicles

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