Characterization of Lactogen Receptor-binding Site 1 of Human Prolactin

Kinet, Sandrina; Goffin, Vincent; Mainfroid, Véronique; Martial, Joseph

doi:10.1074/jbc.271.24.14353

Cited by 38 publications

(33 citation statements)

References 47 publications

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“…As PRL polymers have been long known to display reduced biological activity (for a review, see Ref. 41), we routinely eliminate these multimers from refolded hPRL through a single gel filtration step (25,26,(31)(32)(33)42). As illustrated by SDS-PAGE analysis, this step also eliminates most contaminants.…”

Section: Discussionmentioning

confidence: 99%

“…Based on the three-dimensional model that we established (52), serine 179 is predicted to belong to helix 4, the region of binding site 1 containing the highest number of binding determinants (6). Moreover, the dose-response curves obtained for S179D-hPRL in the various bioassays investigated here are displaced to the right compared with WT hPRL and achieve a maximal response (or supramaximal, see below), two features that fit with alteration of binding site 1 (6,33,42,50). Although S179D-hPRL can be considered as a site 1 analog, this does not necessarily mean that serine 179 is intrinsically involved in an interaction with the PRLR.…”

Section: Fig 6 Cell Cycle Analyses On Human Mammary Tumor Epitheliamentioning

confidence: 99%

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S179D-Human PRL, a Pseudophosphorylated Human PRL Analog, Is an Agonist and Not an Antagonist

et al. 2001

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For many years, our group has been involved in the development of human PRL antagonists. In two recent publications, S179D-human PRL, a human PRL analog designed to mimic a putative S179-phosphorylated human PRL, was reported to be a highly potent antagonist of human PRL-induced proliferation and signaling in rat Nb2 cells. We prepared this analog with the aim of testing it in various bioassays involving the homologous, human PRL receptor. In our hands, S179D-human PRL was able to stimulate 1) the proliferation of rat Nb2 cells and of human mammary tumor epithelial cells (T-47D), 2) transcriptional activation of the lactogenic hormone response element-luciferase reporter gene, and 3) activation of the Janus kinase/signal transducer and activator of transcription and MAPK pathways. Using the previously characterized antagonist G129R-human PRL as a control, we failed to observe any evidence for antagonism of S179D-human PRL toward any of the human PRL-induced effects analyzed, including cell proliferation, transcriptional activation, and signaling. In conclusion, our data argue that S179D-human PRL is an agonist displaying slightly reduced affinity and activity due to local alteration of receptor binding site 1, and that the antagonistic properties previously attributed to S179D-human PRL cannot be confirmed in any of the assays analyzed in this study. (Endocrinology 142: 3950 -3963, 2001)

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Section: Discussionmentioning

confidence: 99%

Section: Fig 6 Cell Cycle Analyses On Human Mammary Tumor Epitheliamentioning

confidence: 99%

S179D-Human PRL, a Pseudophosphorylated Human PRL Analog, Is an Agonist and Not an Antagonist

et al. 2001

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“…Serine 179 is a surprisingly crucial residue to the structure of PRL since its mutation to alanine eliminates biological activity of otherwise unmodified PRL (Chen et al, 1998;Kato et al, 1996). Also, mutational analyses of amino acids on the outside of the helix in this region have shown the region to be important in regulating PRL-receptor interactions (Kinet et al, 1996). Second, even polyclonal antibodies against PRL recognize S179D PRL as somewhat different from unmodified PRL (Chen et al, 1998).…”

Section: Conformation Of S179d Prlmentioning

confidence: 99%

S179D prolactin: Antagonistic agony!

Walker

2007

Molecular and Cellular Endocrinology

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The aims of this review are threefold: First, to collate what is known about the production and activities of phosphorylated prolactin (PRL), the latter largely, but not exclusively, as illustrated through the use of the molecular mimic, S179D PRL; second, to apply this and related knowledge to produce an updated model of prolactin-receptor interactions that may apply to other members of this cytokine super-family; and third, to promote a shift in the current paradigm for the development of clinically important growth antagonists. This third aim explains the title since, based on results with S179D PRL, it is proposed that agents which signal to antagonistic ends may be better therapeutics than pure antagonists -hence antagonistic agony. Since S179D PRL is not a pure antagonist, we have proposed the term selective prolactin receptor modulator (SPeRM) for this and like molecules.

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“…Biochemical Characterization of hPRL Analogs-The content of ␣-helical structure was calculated from circular dichroism spectra obtained as described previously (31,43,45,46). The apparent molecular mass was estimated from the elution volume on gel filtration chromatography (Sephacryl S-100 or S-200 loaded into a C16/70 column; Amersham Biosciences) with respect to the elution volume of standard protein as described elsewhere (46).…”

Section: Methodsmentioning

confidence: 99%

Development of Pure Prolactin Receptor Antagonists

Bernichtein

Kayser

Dillner

et al. 2003

Journal of Biological Chemistry

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Prolactin (PRL) promotes tumor growth in various experimental models and leads to prostate hyperplasia and mammary neoplasia in PRL transgenic mice. Increasing experimental evidence argues for the involvement of autocrine PRL in this process. PRL receptor antagonists have been developed to counteract these undesired proliferative actions of PRL. However, all forms of PRL receptor antagonists obtained to date exhibit partial agonism, preventing their therapeutic use as full antagonists. In the present study, we describe the development of new human PRL antagonists devoid of agonistic properties and therefore able to act as pure antagonists. This was demonstrated using several in vitro bioassays, including highly sensitive assays able to detect extremely low levels of receptor activation. These new compounds also act as pure antagonists in vivo, as assessed by analyzing their ability to competitively inhibit PRL-triggered signaling cascades in various target tissues (liver, mammary gland, and prostate). Finally, by using transgenic mice expressing PRL specifically in the prostate, which exhibit constitutively activated signaling cascades paralleling hyperplasia, we show that these new PRL analogs are able to completely revert PRL-activated events. These second generation human PRL antagonists are good candidates to be used as inhibitors of growth-promoting actions of PRL.

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Characterization of Lactogen Receptor-binding Site 1 of Human Prolactin

Cited by 38 publications

References 47 publications

S179D-Human PRL, a Pseudophosphorylated Human PRL Analog, Is an Agonist and Not an Antagonist

S179D-Human PRL, a Pseudophosphorylated Human PRL Analog, Is an Agonist and Not an Antagonist

S179D prolactin: Antagonistic agony!

Development of Pure Prolactin Receptor Antagonists

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