Isolation and Characterization of Hypothalamic Peptide Hormones

Burgus, Roger; Amoss, Max S.; Brazeau, Paul; Brown, Marvin R.; Ling, Nicholas; Rivier, Catherine; Rivier, Jean; Vale, Wylie; Villarreal, Jose

doi:10.1007/978-1-4684-2598-7_19

Cited by 10 publications

(4 citation statements)

References 44 publications

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“…Gradients were used to determine proper solvent systems for individual compounds but isocratic solvent mixtures gave larger separation factors with closely related compounds. 23 Each analogue (40 Mg) was injected in solution in the solvent mixture used for developing (see legend of Table II for further details).…”

Section: Methodsmentioning

confidence: 99%

Neurotensin analogs. Structure-activity relationships

Rivier¹,

Lazarus²,

Perrin³

et al. 1977

J. Med. Chem.

103

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A series of neurotensin (NT) analogues in which each amino acid has been successively replaced by its D isomer, as well as analogues involving modifications at positions 3 and 11 and a cyclic compound [Cys2,13]-NT, has been synthesized by solid-phase methodology. After purification by conventional techniques the compounds were characterized by thin-layer chromatography, amino acid analysis, and optical rotation. Further characterization of the analogues by high-pressure liquid chromatography demonstrates the high resolving power of this new method. Each analogue was studied for its ability to induce hypothermia in cold-exposed rate (4 degrees C) in vivo and to bind to mast cells in vitro. Although close correlation in potencies was not found for all the analogues tested in both assay systems, they substantiate the basic observation that substitutions in positions 1-9 produced active peptides whereas modification of residues 10-13 considerably decreased biological response in vitro and in vivo. One exception is the higher potency of [D-Phe11]-NT and [D-Tyr11]-NT in vivo. The differences between the efficacies of these analogues in vivo and in vitro are discussed.

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Section: Methodsmentioning

confidence: 99%

Neurotensin analogs. Structure-activity relationships

Rivier¹,

Lazarus²,

Perrin³

et al. 1977

J. Med. Chem.

103

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“…With the availability of synthetic oCRF and antibodies raised against it, various aspects of oCRF's distribution and of its hypophyseal, vascular, and neural actions have been investigated (9)(10)(11)(12)(13)(14)(15)(16)(17)(18), and immunoneutralization experiments in the rat with anti-oCRF antisera have demonstrated that this or related peptides are indeed involved in the physiologic regulation of ACTH secretion (19). As oCRF was purified from a side fraction (5) containing only a small percentage of the total ACTH-releasing activity in ovine hypothalamic extracts (20), it was necessary to establish the structure of the predominant CRF in fresh tissue extracts. Because of its importance as an experimental animal and because its CRF differed from oCRF with respect to immunologic and chromatographic behavior, the rat was chosen as a source of CRF.…”

mentioning

confidence: 99%

Characterization of rat hypothalamic corticotropin-releasing factor.

Rivier¹,

Spiess²,

Vale³

1983

Proc. Natl. Acad. Sci. U.S.A.

281

129

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A polypeptide was purified from rat hypothalamic extracts on the basis of its high intrinsic activity to release corticotropin (ACTH) from cultured rat anterior pituitary cells and its immunoactivity in a radioimmunoassay directed against the NH2 terminus (residues 4-20) of ovine hypothalamic corticotropin-releasing factor (CRF). Based on Edman degradation, peptide mapping, and amino acid analysis, the primary structure of this rat CRF was established to be: H-Ser-Glu-Glu-Pro-Pro-IleSer-Leu-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Glu-Val-Leu-GluMet-Ala-Arg-Ala-Glu-Gln-Leu-Ala-Gln-Gln-Ala-His-Ser-Asn-ArgLys-Leu-Met-Glu-Ile-Ile-NH2. The hypophysiotropic potency of synthetic rat CRF did not deviate significantly from the potencies of the isolated native peptide or of synthetic ovine CRF. The close structural relationship between rat and ovine hypothalamic CRF is indicated by an 83% sequence homology.Harris proposed that the neuroregulation of adrenocorticotropic hormone (ACTH; corticotropin) may be mediated by a substance later termed corticotropin-releasing factor (CRF), which reaches the adenohypophysis by the hypothalamic-hypophysial portal system (1). Early experimental observations by Guillemin and Rosenberg (2), and Saffran and Schally (3), supported the presence of such factors in the hypothalamus that would increase the rate of ACTH secretion by the pituitary gland incubated in vitro or maintained in organ culture. More than 25 years later, ovine CRF (oCRF) was isolated (4, 5), characterized (5, 6), and synthesized (7). Subsequently the amino acid sequence of the protein precursor of oCRF was deduced from the cDNA (8). With the availability of synthetic oCRF and antibodies raised against it, various aspects of oCRF's distribution and of its hypophyseal, vascular, and neural actions have been investigated (9)(10)(11)(12)(13)(14)(15)(16)(17)(18), and immunoneutralization experiments in the rat with anti-oCRF antisera have demonstrated that this or related peptides are indeed involved in the physiologic regulation of ACTH secretion (19). As oCRF was purified from a side fraction (5) containing only a small percentage of the total ACTH-releasing activity in ovine hypothalamic extracts (20), it was necessary to establish the structure of the predominant CRF in fresh tissue extracts. Because of its importance as an experimental animal and because its CRF differed from oCRF with respect to immunologic and chromatographic behavior, the rat was chosen as a source of CRF. We report here the isolation, characterization, and total synthesis of rat CRF (rCRF), a 41-residue peptide belonging to the family that includes oCRF, sauvagine (21), and urotensin 1 (22). In view of the fact that the latter three peptides have a spectrum of biological activities on the cardiovascular and central nervous systems of the mammal with differences in relative potencies (23-25), it was furthermore important to characterize CRF in the species in which its physiological functions have been most thoroughly investigated. EXPERIMENTAL PROCED...

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“…It has recently been shown that reversed-phase liquid chromatography can be used for both the extraction and complete purification of peptides (Bennett et al, 1980a,b). One of the most widely used supports in the RP-HPLC of peptides has been octadecylsilylsilica (ODS-silica) (Burgus & Rivier, 1976; Bennett et al, 1977;Hancock et al, 1978). ODS-silica has a high affinity for peptides and has been used successfully to extract peptides from whole plasma (Bennett et al, 1977) and from tissue homogenates (Bennett et al, 1978).…”

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confidence: 99%