Discovery of the improved antagonistic prolactin variants by library screening

Liu, Yun; Gong, Wenbin; Breinholt, Jens; Nørskov-Lauritsen, Leif; Zhang, Jinchao; Ma, Qinhong; Chen, Jianhe; Panina, Svetlana; Guo, Wei; Li, Tengkun; Zhang, Jingyuan; Kong, Meng; Liu, Zibing; Mao, Jingjing; Christensen, Leif; Hu, Sean; Wang, Lingyun

doi:10.1093/protein/gzr047

Cited by 10 publications

(11 citation statements)

References 16 publications

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“…It is important to note that some women in the low and high quartiles did not have non-synonymous genetic variation in ZnT2, thus variation in other genes are possible modifiers as well. Because PRLR, through activation of the Jak2/Stat5 signaling pathway, increases ZnT2 expression and Zn secretion [38], and genetic variation in PRLR affects milk production traits in production animals [39], we tested the hypothesis that genetic variation in PRLR may underlie the abnormally low or high milk [Zn] in women without variants in ZnT2. Three missense SNPs have been reported in the PRLR extracellular domain; the 'gain-of-function' variants I 100 V and I 146 L, and the 'loss-of-function' variant H 188 R [21][22][23].…”

Section: Alternative Modifiers Of Milk [Zn]mentioning

confidence: 99%

Exome Sequencing of SLC30A2 Identifies Novel Loss- and Gain-of-Function Variants Associated with Breast Cell Dysfunction

Alam

Hennigar

Gallagher

et al. 2015

J Mammary Gland Biol Neoplasia

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The zinc (Zn) transporter ZnT2 (SLC30A2) is expressed in specialized secretory cells including breast, pancreas and prostate, and imports Zn into mitochondria and vesicles. Mutations in SLC30A2 substantially reduce milk Zn concentration ([Zn]) and cause severe Zn deficiency in exclusively breastfed infants. Recent studies show that ZnT2-null mice have low milk [Zn], in addition to profound defects in mammary gland function during lactation. Here, we used breast milk [Zn] to identify novel non-synonymous ZnT2 variants in a population of lactating women. We also asked whether specific variants induce disturbances in intracellular Zn management or cause cellular dysfunction in mammary epithelial cells. Healthy, breastfeeding women were stratified into quartiles by milk [Zn] and exonic sequencing of SLC30A2 was performed. We found that 36% of women tested carried non-synonymous ZnT2 variants, all of whom had milk Zn levels that were distinctly above or below those in women without variants. We identified 12 novel heterozygous variants. Two variants (D(103)E and T(288)S) were identified with high frequency (9 and 16%, respectively) and expression of T(288)S was associated with a known hallmark of breast dysfunction (elevated milk sodium/potassium ratio). Select variants (A(28)D, K(66)N, Q(71)H, D(103)E, A(105)P, Q(137)H, T(288)S and T(312)K) were characterized in vitro. Compared with wild-type ZnT2, these variants were inappropriately localized, and most resulted in either 'loss-of-function' or 'gain-of-function', and altered sub-cellular Zn pools, Zn secretion, and cell cycle check-points. Our study indicates that SLC30A2 variants are common in this population, dysregulate Zn management and can lead to breast cell dysfunction. This suggests that genetic variation in ZnT2 could be an important modifier of infant growth/development and reproductive health/disease. Importantly, milk [Zn] level may serve as a bio-reporter of breast function during lactation.

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Section: Alternative Modifiers Of Milk [Zn]mentioning

confidence: 99%

Exome Sequencing of SLC30A2 Identifies Novel Loss- and Gain-of-Function Variants Associated with Breast Cell Dysfunction

Alam

Hennigar

Gallagher

et al. 2015

J Mammary Gland Biol Neoplasia

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“…Studies have shown that Prl G129R competes with exogenous Prl when added in >10-fold molar excess to compensate for its lower affinity for the PrlR [48, 49]. In the present study we used a protein with increased affinity to block the receptor, Prl S33A, Q73L, G129R, K190R [54]. …”

Section: Methodsmentioning

confidence: 99%

Stimulation of prolactin receptor induces STAT-5 phosphorylation and cellular invasion in glioblastoma multiforme

Alkharusi

Landázuri

et al. 2016

Oncotarget

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Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumor in humans and is characterized with poor outcome. In this study, we investigated components of prolactin (Prl) system in cell models of GBM and in histological tissue sections obtained from GBM patients. Expression of Prolactin receptor (PrlR) was detected at high levels in U251-MG, at low levels in U87-MG and barely detectable in U373 cell lines and in 66% of brain tumor tissues from 32 GBM patients by immunohistochemical technique. In addition, stimulation of U251-MG and U87-MG cells but not U373 with Prl resulted in increased STAT5 phosphorylation and only in U251-MG cells with increased cellular invasion. Furthermore, STAT5 phosphorylation and cellular invasion induced in Prl stimulated cells were significantly reduced by using a Prl receptor antagonist that consists of Prl with four amino acid replacements. We conclude that Prl receptor is expressed at different levels in the majority of GBM tumors and that blocking of PrlR in U251-MG cells significantly reduce cellular invasion.

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“…This modified hPRL and an additional mutant in which the N-terminal nine amino acids were deleted (D1-9-G129R have provided a wealth of information on the receptor binding theory for PRL, reviewed in Goffin et al (2005). Recently, a separate group has identified mutations in binding site 1 that improved the potency of G129R hPRL (Liu et al 2011), the effect of combining these site 1 mutations with D1-9-G129R in hPRL has not yet been tested.…”

Section: Signal Transductionmentioning

confidence: 99%