Abstract
Bullock median-eminence tissue was used as a source of luteinizing-hormone-releasing factor for small-scale experiments to explore methods for its isolation. The presence of luteinizing-hormone-releasing factor was detected by the ovulation response in rabbits after intrapituitary infusion of the extract. Gel filtration was found to be suitable for the purification of these extracts. The releasing factor appeared to be a basic peptide of molecular weight in the range 1200–2500. On a larger scale, an extract of hypothalamic tissue from sheep was used to establish a multi-stage isolation procedure that resulted in a 200000-fold purification of luteinizing-hormone-releasing factor. After the initial extraction the isolation process consisted of: (1) two cycles of gel filtration; (2) anion-exchange chromatography; (3) gel filtration in a partially organic medium; (4) thin-layer chromatography on cellulose. Stage (3) separated two zones of activity each containing peptides. One of these was purified further by stage (4) to give a preparation that was active at a dose of 6μg. of peptide/animal, although activity diminished seriously during storage. This preparation contained only five or six components, but the small amounts of peptides obtained at this stage of purity were insufficient for full characterization.
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- CAMPBELL H. J., FEUER G., HARRIS G. W. THE EFFECT OF INTRAPITUITARY INFUSION OF MEDIAN EMINENCE AND OTHER BRAIN EXTRACTS ON ANTERIOR PITUITARY GONADOTROPHIC SECRETION. J Physiol. 1964 Apr;170:474–486. doi: 10.1113/jphysiol.1964.sp007344. [DOI] [PMC free article] [PubMed] [Google Scholar]
- COURRIER R., JUTISZ M., COLONGE A. A PROPOS DE LA PR'ESENCE D'ACTIVIT'ES HORMONALES DE TYPE ANT'EHYPOPHYSAIRE DANS LES EXTRAITS D'HYPOTHALAMUS. C R Hebd Seances Acad Sci. 1963 Dec 16;257:3774–3776. [PubMed] [Google Scholar]
- DHARIWAL A. P., ANTUNES-RODRIGUES J., MCCANN S. M. PURIFICATION OF OVINE LUTEINIZING HORMONE-RELEASING FACTOR BY GEL FILTRATION AND ION EXCHANGE CHROMATOGRAPHY. Proc Soc Exp Biol Med. 1965 Apr;118:999–1003. doi: 10.3181/00379727-118-30028. [DOI] [PubMed] [Google Scholar]
- ELLIOTT D. F., HORTON E. W., LEWIS G. P. The isolation of bradykinin, a plasma kinin from ox blood. Biochem J. 1961 Jan;78:60–65. doi: 10.1042/bj0780060. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Endröczi E., Hilliard J. Luteinizing hormone releasing activity in different parts of rabbit and dog brain. Endocrinology. 1965 Oct;77(4):667–673. doi: 10.1210/endo-77-4-667. [DOI] [PubMed] [Google Scholar]
- GUILLEMIN R., COLONGE A., JUTISZ M., SAKIZ E. [Persistence of the biological activity of the luteinizing hormone after heating to 100 degrees C in an acid medium]. C R Hebd Seances Acad Sci. 1962 Apr 30;254:3258–3260. [PubMed] [Google Scholar]
- GUILLEMIN R., JUTISZ M., SAKIZ E. [The partial purification of an hypothalamic factor (LRF) stimulating the secretion of pituitary luteinizing hormone (LH)]. C R Hebd Seances Acad Sci. 1963 Jan 7;256:504–507. [PubMed] [Google Scholar]
- GUILLEMIN R., SCHALLY A. V., LIPSCOMB H. S., ANDERSEN R. N., LONG J. M. On the presence in hog hypothalamus of 3-corticotropin releasing factor, alpha- and beta-melanocyte stimulating hormones, adrenocorticotropin, lysine-vasopressin and oxytocin. Endocrinology. 1962 Apr;70:471–477. doi: 10.1210/endo-70-4-471. [DOI] [PubMed] [Google Scholar]
- HEILMANN J., BARROLLIER J., WATZKE E. Beitrag zur Aminosäurebestimmung auf Papierchromatogrammen. Hoppe Seylers Z Physiol Chem. 1957;309(4-6):219–220. [PubMed] [Google Scholar]
- Harris G. W., Reed M., Fawcett C. P. Hypothalamic releasing factors and the control of anterior pituitary function. Br Med Bull. 1966 Sep;22(3):266–272. doi: 10.1093/oxfordjournals.bmb.a070485. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- SCHALLY A. V., BOWERS C. Y. PURIFICATION OF LUTEINIZING HORMONE-RELEASING FACTOR FROM BOVINE HYPOTHALAMUS. Endocrinology. 1964 Oct;75:608–614. doi: 10.1210/endo-75-4-608. [DOI] [PubMed] [Google Scholar]
- STOUFFER J. E., LIPSCOMB H. S. Reversibel binding of ACTH to a glass surface. Endocrinology. 1963 Jan;72:91–94. doi: 10.1210/endo-72-1-91. [DOI] [PubMed] [Google Scholar]
- WINNICK T., WINNICK R. E., ACHER R., FROMAGEOT C. Amino acids and peptides of posterior pituitary and hypothalamus tissues. Biochim Biophys Acta. 1955 Dec;18(4):488–494. doi: 10.1016/0006-3002(55)90139-6. [DOI] [PubMed] [Google Scholar]

