scholarly journals Quantitative measurement of allele‐specific protein expression in a diploid yeast hybrid by LC‐MS

2012 ◽  
Vol 8 (1) ◽  
pp. 602 ◽  
Author(s):  
Zia Khan ◽  
Joshua S Bloom ◽  
Sasan Amini ◽  
Mona Singh ◽  
David H Perlman ◽  
...  
Keyword(s):  
Protein Expression ◽  
Quantitative Measurement ◽  
Specific Protein ◽  
Allele Specific ◽  
Yeast Hybrid

scholarly journals Determining Allele-Specific Protein Expression (ASPE) Using a QconCAT-Based Proteomics Method: a Novel Approach to Identify Cis-acting Genetic Variants

2018 ◽  
Author(s):  
Jian Shi ◽  
Xinwen Wang ◽  
Huaijun Zhu ◽  
Hui Jiang ◽  
Danxin Wang ◽  
...  
Keyword(s):  
Protein Expression ◽  
Genetic Variants ◽  
Multiple Reaction Monitoring ◽  
Specific Protein ◽  
Wild Type ◽  
Specific Expression ◽  
Cis Acting ◽  
Regulatory Variants ◽  
Novel Approach ◽  
Allele Specific

AbstractMeasuring allele-specific expression (ASE) is a powerful approach for identifying cis-regulatory genetic variants. Here we developed a novel targeted proteomics method for quantification of allele-specific protein expression (ASPE) based on scheduled high resolution multiple reaction monitoring (sMRM-HR) with a heavy stable isotope-labeled quantitative concatamer (QconCAT) internal protein standard. This strategy was applied to the determination of the ASPE of UGT2B15 in human livers using the common UGT2B15 nonsynonymous variant rs1902023 (i.e. Y85D) as the marker to differentiate expressions from the two alleles. The QconCAT standard contains both the wild type tryptic peptide and the Y85D mutant peptide at a ratio of 1:1 to ensure accurate measurement of the ASPE of UGT2B15. The results from 18 UGT2B15 Y85D heterozygotes revealed that the ratios between wild type Y allele and mutant D allele varied from 0.60 to 1.46, indicating the presence of cis-regulatory variants. In addition, we observed no significant correlations between the ASPE and mRNA ASE of UGT2B15, suggesting the involvement of different cis-acting variants in regulating the transcription and translation processes of the gene. This novel ASPE approach provides a powerful tool for capturing cis-genetic variants involved in post-transcription processes, an important yet understudied area of research.

scholarly journals Determining Allele-Specific Protein Expression (ASPE) Using a Novel Quantitative Concatamer Based Proteomics Method

2018 ◽  
Vol 17 (10) ◽  
pp. 3606-3612 ◽  
Author(s):  
Jian Shi ◽  
Xinwen Wang ◽  
Huaijun Zhu ◽  
Hui Jiang ◽  
Danxin Wang ◽  
...  
Keyword(s):  
Protein Expression ◽  
Specific Protein ◽  
Allele Specific

Response to the Comments on “Determining Allele-Specific Protein Expression (ASPE) Using a Novel Quantitative Concatamer Proteomics Method”

2019 ◽  
Vol 18 (3) ◽  
pp. 1458-1459
Author(s):  
Jian Shi ◽  
Xinwen Wang ◽  
Huaijun Zhu ◽  
Hui Jiang ◽  
Danxin Wang ◽  
...  
Keyword(s):  
Protein Expression ◽  
Specific Protein ◽  
Allele Specific

scholarly journals Parental Allele-Specific Protein Expression in Single Cells In Vivo

2018 ◽  
Author(s):  
Chiu-An Lo ◽  
Brian E. Chen
Keyword(s):  
Protein Expression ◽  
Single Cells ◽  
Specific Protein ◽  
The Other ◽  
Parental Allele ◽  
Allele Specific ◽  
Parent Of Origin ◽  
Ribosomal Protein L13a ◽  
Over Time

AbstractAllelic expression from each parent-of-origin is important as a backup and to ensure that enough protein products of a gene are produced. Thus far, it is not known how each cell throughout a tissue differs in parental allele expression at the level of protein synthesis. Here, we measure the expression of the Ribosomal protein L13a (Rpl13a) from both parental alleles simultaneously in single cells in the living animal. We use genome-edited Drosophila that have a quantitative reporter of protein synthesis inserted into the endogenous Rpl13a locus. We find that individual cells can have large (>10-fold) differences in protein expression between the two parental alleles. Cells can produce protein from only one allele oftentimes, and time-lapse imaging of protein production from each parental allele in each cell showed that the imbalance in expression from one parental allele over the other can invert over time.One sentence summaryParental allele-specific protein expression varies widely across cells and over time.HighlightsWe used genome editing to insert a quantifiable protein translation reporter into the endogenous Ribosomal protein L13a gene and thus track the protein expression of both parental alleles simultaneously in every single cell in the awake animal.Cells can have a large difference in protein expression for one parental allele over the other, and this can invert over time, and can occur in clusters of cells within a tissue.We demonstrate the highly variable nature of heterozygous and homozygous definitions across single cells, and over time.Our study demonstrates a new paradigm that can be used to examine inherited epigenetic control of expression from a specific parent-of-origin allele across single clonal cells from a common progenitor in vivo.

Tissue Distribution and Gender-Specific Protein Expression of Cytochrome P450 in five Mouse Genotypes with a Background of FVB

2018 ◽  
Vol 35 (6) ◽  
Author(s):  
Jiamei M. Chen ◽  
Qisong S. Zhang ◽  
Xiaoyan Y. Li ◽  
Xia Gong ◽  
Yanjiao J. Ruan ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Protein Expression ◽  
Tissue Distribution ◽  
Specific Protein ◽  
And Gender ◽  
Gender Specific

Fluoride exposure inhibits protein expression and enzyme activity in the lung-stage larvae ofAscaris suum

Parasitology ◽  
2006 ◽  
Vol 133 (4) ◽  
pp. 497-508 ◽  
Author(s):  
M. K. ISLAM ◽  
T. MIYOSHI ◽  
M. YAMADA ◽  
M. A. ALIM ◽  
X. HUANG ◽  
...  
Keyword(s):  
Protein Expression ◽  
Expression Profiles ◽  
Immunoblot Analysis ◽  
Specific Protein ◽  
Inorganic Pyrophosphatase ◽  
2D Electrophoresis ◽  
Peptide Mass ◽  
Enzyme Families ◽  
Protein Expression Profiles ◽  
Peptide Mass Spectrometry

Sodium fluoride (NaF) is an anion that has been previously shown to block the moulting process ofAscaris suumlarvae. This study describes moulting and development-specific protein expression profiles ofA. suumlung-stage L3 (AsLL3) following NaF exposure. AsLL3s cultured in the presence or absence of NaF were prepared for protein analysis using two-dimensional (2D) electrophoresis. NaF exposure inhibited at least 22 proteins in AsLL3 compared with moulted larvae (i.e. AsLL4). A further comparison of AsLL4 with those of pre-cultured AsLL3 and NaF-exposed AsLL3 revealed 8 stage-specifically and 4 over-expressed proteins. Immunoblot analysis revealed an inhibition by NaF of 19 immunoreactive proteins. Enzyme assay and immunochemical data showed an inhibition of the moulting-specific inorganic pyrophosphatase activity by 41% and a decreased expression in NaF-treated larvae, indicating its significance in the moulting process. A protein spot associated with NaF inhibition was isolated and identified by peptide mass spectrometry and bioinformatics approaches to be a member of 3–hydroxyacyl–CoA dehydrogenase/short-chain dehydrogenase enzyme families. These results have implications for the identification of proteins specific to the moulting process as potential chemotherapeutic targets.

Sign in / Sign up

Export Citation Format

Share Document