Proteomic analysis of porcine endometrial tissue during peri-implantation period reveals altered protein abundance

Recent evidence points to a role for peroxisome proliferator-activated receptors (PPARs) δ and γ in embryo implantation and survival. In this study, we report the porcine PPARδ complete coding sequence and mRNA abundance of PPARδ, PPARγ1 and γ2, angiopoietin-like protein 4 (ANGPTL4) and adipocyte determination and differentiation-dependent factor 1 (ADD1) genes in the pregnant sow endometrium. Real-time PCR analysis was used to study the effect of parity (Yorkshire-Landrace multiparous (YL) and nulliparous (YLn)), site of endometrial tissue sampling (between and at embryo attachment sites) in crossbred Duroc×Yorkshire-Landrace (DYL) sows and stages of pregnancy (non-pregnant, day 15 and day 25 after mating) in Meishan-Landrace (ML) on mRNA levels. Parity effects were observed for PPARδ, ANGPTL4, and ADD1, with higher mRNA levels in YL than YLn sows. In DYL sows, lower mRNA levels were present at attachment sites compared to between attachment sites for PPARδ, PPARγ1, and ANGPTL4. Finally, day 15 pregnant ML sows had lower PPARδ mRNA levels compared to day 15 cycling ML sows. A significant increase of PPARγ1 mRNA levels was found on day 25 pregnant ML and DYL sows relative to day 15 ML or DYL pregnant sows. PPARδ and γ immunostaining was detected in endometrial tissue of day 15 cycling sows, day 15 and 25 pregnant sows and epithelial cells of day 25 embryos. Collectively, our results suggest a role for PPARδ, PPARγ1, and ANGPTL4, but not PPARγ2, during the peri-implantation period in pregnant sows.

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Sucrose Metabolism in Haloarchaea: Reassessment Using Genomics, Proteomics, and Metagenomics

Applied and Environmental Microbiology ◽

10.1128/aem.02935-18 ◽

2019 ◽

Vol 85 (6) ◽

Cited By ~ 2

Author(s):

Timothy J. Williams ◽

Michelle A. Allen ◽

Yan Liao ◽

Mark J. Raftery ◽

Ricardo Cavicchioli

Keyword(s):

Proteomic Analysis ◽

Sucrose Metabolism ◽

Metagenomic Data ◽

Protein Abundance ◽

Metabolic Capacity ◽

Sucrose Hydrolysis ◽

Content Type ◽

Fructose Metabolism ◽

Novel Proteins ◽

Related Proteins

ABSTRACTThe canonical pathway for sucrose metabolism in haloarchaea utilizes a modified Embden-Meyerhof-Parnas pathway (EMP), in which ketohexokinase and 1-phosphofructokinase phosphorylate fructose released from sucrose hydrolysis. However, our survey of haloarchaeal genomes determined that ketohexokinase and 1-phosphofructokinase genes were not present in all species known to utilize fructose and sucrose, thereby indicating that alternative mechanisms exist for fructose metabolism. A fructokinase gene was identified in the majority of fructose- and sucrose-utilizing species, whereas only a small number possessed a ketohexokinase gene. Analysis of a range of hypersaline metagenomes revealed that haloarchaeal fructokinase genes were far more abundant (37 times) than haloarchaeal ketohexokinase genes. We used proteomic analysis ofHalohasta litchfieldiae(which encodes fructokinase) and identified changes in protein abundance that relate to growth on sucrose. Proteins inferred to be involved in sucrose metabolism included fructokinase, a carbohydrate primary transporter, a putative sucrose hydrolase, and two uncharacterized carbohydrate-related proteins encoded in the same gene cluster as fructokinase and the transporter. Homologs of these proteins were present in the genomes of all haloarchaea that use sugars for growth. Enzymes involved in the semiphosphorylative Entner-Doudoroff pathway also had higher abundances in sucrose-grownH. litchfieldiaecells, consistent with this pathway functioning in the catabolism of the glucose moiety of sucrose. The study revises the current understanding of fundamental pathways for sugar utilization in haloarchaea and proposes alternatives to the modified EMP pathway used by haloarchaea for sucrose and fructose utilization.IMPORTANCEOur ability to infer the function that microorganisms perform in the environment is predicated on assumptions about metabolic capacity. When genomic or metagenomic data are used, metabolic capacity is inferred from genetic potential. Here, we investigate the pathways by which haloarchaea utilize sucrose. The canonical haloarchaeal pathway for fructose metabolism involving ketohexokinase occurs only in a small proportion of haloarchaeal genomes and is underrepresented in metagenomes. Instead, fructokinase genes are present in the majority of genomes/metagenomes. In addition to genomic and metagenomic analyses, we used proteomic analysis ofHalohasta litchfieldiae(which encodes fructokinase but lacks ketohexokinase) and identified changes in protein abundance that related to growth on sucrose. In this way, we identified novel proteins implicated in sucrose metabolism in haloarchaea, comprising a transporter and various catabolic enzymes (including proteins that are annotated as hypothetical).

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Short-term high-glucose treatment decreased abundance of Orai1 protein through posttranslational mechanisms in rat mesangial cells

AJP Renal Physiology ◽

10.1152/ajprenal.00513.2017 ◽

2018 ◽

Vol 314 (5) ◽

pp. F855-F863 ◽

Cited By ~ 2

Author(s):

Hui Jiang ◽

Shubiao Zou ◽

Sarika Chaudhari ◽

Rong Ma

Keyword(s):

High Glucose ◽

Mesangial Cells ◽

Protein Synthesis Inhibitor ◽

Protein Abundance ◽

Oxygen Species ◽

Short Term ◽

Synthesis Inhibitor ◽

Altered Protein ◽

Short Period ◽

Ubiquitin Proteasome

The short-term effect of high-glucose (HG) treatment on store-operated Ca2+ entry in mesangial cells (MCs) is not well-known. The aim of the present study was to determine whether and how HG treatment for a short period altered protein abundance of Orai1, the channel mediating store-operated Ca2+ entry in MCs. Rat and human MCs were exposed to HG (25 mM) for 2, 4, 8, and 24 h, and the abundance of Orai1 protein was significantly decreased at the time points of 8 and 16 h. Consistently, HG treatment for 8 h significantly reduced store-operated Ca2+ entry in rat MCs. However, HG treatment for the same time periods did not alter the levels of Orai1 transcript. Cycloheximide, a protein synthesis inhibitor, did not affect the HG-induced decrease of Orai1 protein, suggesting a posttranslational mechanism was involved. However, the HG effect on Orai1 protein was significantly attenuated by MG132 (a ubiquitin-proteasome inhibitor) and NH4Cl (a lysosomal pathway inhibitor). Furthermore, HG treatment for 8 h stimulated ubiquitination of Orai1 protein. We further found that polyethylene glycol-catalase, an antioxidant, significantly blunted the HG-induced reduction of Orai1 protein. In support of involvement of reactive oxygen species in the HG effects, hydrogen peroxide (H2O2) itself significantly decreased abundance of Orai1 protein and increased the level of ubiquitinated Orai1. Taken together, these results suggest that a short-term HG treatment decreased abundance of Orai1 protein in MCs by promoting the protein degradation through the ubiquitination-proteasome and –lysosome mechanisms. This HG-stimulated posttranslational mechanism was mediated by H2O2.

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164 Proteomic analysis of liver mitochondria proteins: evidence for altered protein thiol redox status after chronic alcohol consumption

Hepatology ◽

10.1016/s0270-9139(03)80207-0 ◽

2003 ◽

Vol 38 ◽

pp. 234-235

Author(s):

S BAILEY ◽

A VENKATRAMAN ◽

A LANDAR ◽

A DAVIS ◽

E OULASOVA ◽

...

Keyword(s):

Alcohol Consumption ◽

Proteomic Analysis ◽

Liver Mitochondria ◽

Redox Status ◽

Chronic Alcohol ◽

Protein Thiol ◽

Chronic Alcohol Consumption ◽

Altered Protein ◽

Thiol Redox

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Proteomic analysis of cell cycle progression in asynchronous cultures, including mitotic subphases, using PRIMMUS

eLife ◽

10.7554/elife.27574 ◽

2017 ◽

Vol 6 ◽

Cited By ~ 27

Author(s):

Tony Ly ◽

Arlene Whigham ◽

Rosemary Clarke ◽

Alejandro J Brenes-Murillo ◽

Brett Estes ◽

...

Keyword(s):

Gene Expression ◽

Mass Spectrometry ◽

Proteomic Analysis ◽

Cell Cycle Progression ◽

Protein Abundance ◽

Phosphorylation Sites ◽

Mitotic Progression ◽

Cycle Progression ◽

Post Translational Modifications ◽

Temporal Regulation

The temporal regulation of protein abundance and post-translational modifications is a key feature of cell division. Recently, we analysed gene expression and protein abundance changes during interphase under minimally perturbed conditions (Ly et al., 2014, 2015). Here, we show that by using specific intracellular immunolabelling protocols, FACS separation of interphase and mitotic cells, including mitotic subphases, can be combined with proteomic analysis by mass spectrometry. Using this PRIMMUS (PRoteomic analysis of Intracellular iMMUnolabelled cell Subsets) approach, we now compare protein abundance and phosphorylation changes in interphase and mitotic fractions from asynchronously growing human cells. We identify a set of 115 phosphorylation sites increased during G2, termed ‘early risers’. This set includes phosphorylation of S738 on TPX2, which we show is important for TPX2 function and mitotic progression. Further, we use PRIMMUS to provide the first a proteome-wide analysis of protein abundance remodeling between prophase, prometaphase and anaphase.

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Electromagnetic Field (EMF) Radiation Alters Estrogen Release from the Pig Myometrium during the Peri-Implantation Period

International Journal of Molecular Sciences ◽

10.3390/ijms22062920 ◽

2021 ◽

Vol 22 (6) ◽

pp. 2920

Author(s):

Ewa Monika Drzewiecka ◽

Wiktoria Kozlowska ◽

Agata Zmijewska ◽

Pawel Jozef Wydorski ◽

Anita Franczak

Keyword(s):

Electromagnetic Field ◽

Estrogenic Activity ◽

Transcript Abundance ◽

Hydroxysteroid Dehydrogenase ◽

Protein Abundance ◽

Mrna Transcript ◽

Cytochrome P450 Aromatase ◽

Type 3 ◽

Estradiol 17Β ◽

Implantation Period

An electromagnetic field (EMF) may affect the functions of uterine tissues. This study hypothesized that EMF changes the estrogenic activity of pig myometrium during the peri-implantation period. Tissue was collected on days 15–16 of the gestation and incubated in the presence of EMF (50 and 120 Hz, 2 and 4 h). The cytochrome P450 aromatase type 3 (CYP19A3) and hydroxysteroid 17β dehydrogenase type 4 (HSD17B4) mRNA transcript abundance, cytochrome P450arom (aromatase), and 17β hydroxysteroid dehydrogenase 17βHSD) protein abundance and estrone (E1) and estradiol-17β (E2) release were examined using Real-Time PCR, Western blot and radioimmunoassay. Selected myometrial slices were treated with progesterone (P4) to determine whether it functions as a protector against EMF. CYP19A3 mRNA transcript abundance in slices treated with EMF was less at 50 Hz (2 h) and greater at 120 Hz (2 and 4 h). HSD17B4 mRNA transcript was greater in slices treated with EMF at 120 Hz (2 h). Progesterone diminished EMF-related effects on CYP19A3 and HSD17B4. When P4 was added, EMF had suppressive (50 and 120 Hz, 2 h) or enhancing (50 Hz, 4 h) effects on aromatase abundance. The E1 release was lower after 4 h of EMF treatment at 50 Hz and P4 did not protect myometrial E1 release. In conclusion, EMF alters the synthesis and release of E1 and did not affect E2 release in the myometrium during the peri-implantation period.

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Altered protein expression and protein nitration pattern during d-galactosamine-induced cell death in human hepatocytes: a proteomic analysis

Liver International ◽

10.1111/j.1478-3231.2005.01172.x ◽

2005 ◽

Vol 25 (6) ◽

pp. 1259-1269 ◽

Cited By ~ 17

Author(s):

Antonio Rodriguez-Ariza ◽

Laura M. Lopez-Sanchez ◽

Raul Gonzalez ◽

Fernando J. Corrales ◽

Pedro Lopez ◽

...

Keyword(s):

Cell Death ◽

Protein Expression ◽

Proteomic Analysis ◽

Human Hepatocytes ◽

Protein Nitration ◽

Altered Protein

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Proteomic analysis of cell cycle progression in asynchronous cultures, including mitotic subphases, using PRIMMUS

10.1101/125831 ◽

2017 ◽

Author(s):

Tony Ly ◽

Arlene Whigham ◽

Rosemary Clarke ◽

Alejandro Brenes-Murillo ◽

Brett Estes ◽

...

Keyword(s):

Gene Expression ◽

Mass Spectrometry ◽

Proteomic Analysis ◽

Cell Cycle Progression ◽

Protein Abundance ◽

Phosphorylation Sites ◽

Mitotic Progression ◽

Cycle Progression ◽

Post Translational Modifications ◽

Temporal Regulation

AbstractThe temporal regulation of protein abundance and post-translational modifications is a key feature of cell division. Recently, we analysed gene expression and protein abundance changes during interphase under minimally perturbed conditions (Ly et al. 2014; Ly et al. 2015). Here we show that by using specific intracellular immunolabeling protocols, FACS separation of interphase and mitotic cells, including mitotic subphases, can be combined with proteomic analysis by mass spectrometry. Using this PRIMMUS (PRoteomic analysis of Intracellular iMMUnolabeled cell Subsets) approach, we now compare protein abundance and phosphorylation changes in interphase and mitotic fractions from asynchronously growing human cells. We identify a set of 115 phosphorylation sites increased during G2, which we term ‘early risers’. This set includes phosphorylation of S738 on TPX2, which we show is important for TPX2 function and mitotic progression. Further, we use PRIMMUS to provide a proteome-wide analysis of protein abundance remodeling between prophase, prometaphase and anaphase.

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