ArticleAnalytical chemistry2012
Sample prefractionation for mass spectrometry quantification of low-abundance membrane proteins.
Article in Analytical chemistry, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed, 26 citations in OpenAlex.
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- [Recent advances in microchip liquid chromatography].Se pu = Chinese journal of chromatography · 2021Article
- Multiplexed Liquid Chromatography-Multiple Reaction Monitoring Mass Spectrometry Quantification of Cancer Signaling Proteins.Methods in molecular biology (Clifton, N.J.) · 2017Article
- Retinal Hypercholesterolemia Triggers Cholesterol Accumulation and Esterification in Photoreceptor Cells.The Journal of biological chemistry · 2016Article
- Quantification of Borrelia burgdorferi Membrane Proteins in Human Serum: A New Concept for Detection of Bacterial Infection.Analytical chemistry · 2015Article
- Pathways of cholesterol homeostasis in mouse retina responsive to dietary and pharmacologic treatments.Journal of lipid research · 2015Article
- Retinal and nonocular abnormalities in Cyp27a1(-/-)Cyp46a1(-/-) mice with dysfunctional metabolism of cholesterol.The American journal of pathology · 2014Article
- Cholesterol in the retina: the best is yet to come.Progress in retinal and eye research · 2014Review
- Pharmacologic stimulation of cytochrome P450 46A1 and cerebral cholesterol turnover in mice.The Journal of biological chemistry · 2014Article
- Mass spectrometry quantification of PICALM and AP180 in human frontal cortex and neural retina.Analytical biochemistry · 2013Article
- Mass spectrometry assessment of ubiquitin carboxyl-terminal hydrolase L1 partitioning between soluble and particulate brain homogenate fractions.Analytical chemistry · 2013Article
- Mass spectrometry of membrane proteins: a focus on aquaporins.Biochemistry · 2013Review
- Posttranslational modification by an isolevuglandin diminishes activity of the mitochondrial cytochrome P450 27A1.Journal of lipid research · 2013Article
- 15N-labeled full-length apolipoprotein E4 as an internal standard for mass spectrometry quantification of apolipoprotein E isoforms.Analytical chemistry · 2012Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
Use of stable isotope-labeled full-length proteins as an internal standard prior to multiple reaction monitoring (MRM) analysis enables prefractionation of the target proteins and quantification of those low-abundance proteins, which cannot be reached without biological sample enrichment. In terms of membrane proteins, this benefit can be used if a sample processing workflow allows entire solubilization of membrane proteins. We have developed a universal workflow for sample processing and enrichment by optimizing washing and solubilization conditions and implementing sample fractionation by Whole Gel Eluter. The optimized protocol was applied to various membrane-bound cytochromes P450 (CYPs) and their electron transferring protein partners, cytochrome P450 reductase (CPR), ferredoxin reductase (FdR), and ferredoxin (Fdx), all important proteins for cholesterol elimination from different organs. Both, weakly associated (CPR and FdR) and tightly associated (CYP7B1, CYP11A1, CYP27A1, and CYP46A1) membrane proteins were quantified. Measurements were performed on three human tissues (temporal lobe of the brain, retina, and retinal pigment epithelium) obtained from multiple donors. The biological implications of our quantitative measurements are also discussed.
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