A Survey of Membrane Proteins in Human Serum

Dũng, Nguyễn Tiến; Van, Phan

doi:10.4137/pri.s9374

Cited by 4 publications

(5 citation statements)

References 27 publications

(27 reference statements)

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“…Introducing serum sample collection in clinical practice from a population health perspective is more convenient and accessible compared to RNA samples. Thus, serum samples may be a more favorable material used as a biomarker of fatty acid metabolism disturbances in further clinical practice [ 17 , 25 ].…”

Section: Discussionmentioning

confidence: 99%

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The Role of GPR120 Receptor in Essential Fatty Acids Metabolism in Schizophrenia

et al. 2020

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A growing body of evidence confirms abnormal fatty acid (FAs) metabolism in the pathophysiology of schizophrenia. Omega-3 polyunsaturated fatty acids (PUFAs) are endogenous ligands of the G protein-coupled receptors, which have anti-inflammatory properties and are a therapeutic target in many diseases. No clinical studies are concerned with the role of the GPR120 signaling pathway in schizophrenia. The aim of the study was to determine the differences in PUFA nutritional status and metabolism between patients with schizophrenia (SZ group) and healthy individuals (HC group). The study included 80 participants (40 in the SZ group, 40 in the HC group). There were no differences in serum GPR120 and PUFA concentrations and PUFA intake between the examined groups. In the HC group, there was a relationship between FAs in serum and GPR120 concentration (p < 0.05): α-linolenic acid (ALA) (R = −0.46), docosahexaenoic acid (DHA) (R = −0.54), omega-3 PUFAs (R = −0.41), arachidonic acid (AA) (R = −0.44). In the SZ group, FA serum concentration was not related to GPR120 (p > 0.05). In the HC group, ALA and DHA serum concentrations were independently associated with GPR120 (p < 0.05) in the model adjusted for eicosapentaenoic acid (EPA) and accounted for 38.59% of GPR120 variability (p < 0.05). Our results indicate different metabolisms of FAs in schizophrenia. It is possible that the diminished anti-inflammatory response could be a component connecting GPR120 insensitivity with schizophrenia.

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Section: Discussionmentioning

confidence: 99%

“…The number of studies focusing on membrane proteins in serum is still limited. Nevertheless, this type of analysis has the potential to diagnose and/or treat diseases in further clinical practice [17].…”

Section: Gpr120 Assaymentioning

confidence: 99%

The Role of GPR120 Receptor in Essential Fatty Acids Metabolism in Schizophrenia

et al. 2020

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“…Considering the previously identified complement of membranous material and membrane protein in serum and plasma [ 27 , 28 ], ultracentrifugation was carried out according to Churchward et al, (2005) ( supplementary methods ) [ 29 ]. 2 mL of thawed serum with an added 1× conc of PI was centrifuged at 146,542× g for three hours at 4 °C using a Beckman Coulter Optima L-100 XP ultracentrifuge; the separate supernatant and pellet fractions were collected.…”

Section: Methodsmentioning

confidence: 99%

A Routine ‘Top-Down’ Approach to Analysis of the Human Serum Proteome

2017

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Serum provides a rich source of potential biomarker proteoforms. One of the major obstacles in analysing serum proteomes is detecting lower abundance proteins owing to the presence of hyper-abundant species (e.g., serum albumin and immunoglobulins). Although depletion methods have been used to address this, these can lead to the concomitant removal of non-targeted protein species, and thus raise issues of specificity, reproducibility, and the capacity for meaningful quantitative analyses. Altering the native stoichiometry of the proteome components may thus yield a more complex series of issues than dealing directly with the inherent complexity of the sample. Hence, here we targeted method refinements so as to ensure optimum resolution of serum proteomes via a top down two-dimensional gel electrophoresis (2DE) approach that enables the routine assessment of proteoforms and is fully compatible with subsequent mass spectrometric analyses. Testing included various fractionation and non-fractionation approaches. The data show that resolving 500 µg protein on 17 cm 3–10 non-linear immobilised pH gradient strips in the first dimension followed by second dimension resolution on 7–20% gradient gels with a combination of lithium dodecyl sulfate (LDS) and sodium dodecyl sulfate (SDS) detergents markedly improves the resolution and detection of proteoforms in serum. In addition, well established third dimension electrophoretic separations in combination with deep imaging further contributed to the best available resolution, detection, and thus quantitative top-down analysis of serum proteomes.

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“…3A). Since membrane proteins have been reported to be secreted from cells as exosomes to enter circulation [17], we next examined the level of MAS-1, the receptor for Ang-(1-7), in the blood from the subjects.…”

Section: Upregulation Of Mas-1 But Not Ang-(1-7) In Type 2 Diabetes In Humansmentioning

confidence: 99%

“…It is interesting to note that membrane proteins MAS-1 and CFTR are detectable in human blood samples. Since it is known that membrane proteins can be released with exosomes into extracellular fluid [17], we suppose the detected MAS-1 and CFTR proteins are secreted proteins into the circulation system. Importantly, using the hCFTR overexpression CHO cells together with patch-clamp, we have shown positive correlation between ELISA-detected the amount of proteins that are released into the extracellular solution and functional expression of the protein in the cell membrane (supplementary Figure1), validating the ELISA examination of human blood samples to reflex the proteins' in situ membrane expression.…”

Section: Upregulation Of Cftr Correlated With Mas-1 In Type 2 Diabetes In Humansmentioning

confidence: 99%

Angiotensin(1–7) activates MAS-1 and upregulates CFTR to promote insulin secretion in pancreatic β-cells: the association with type 2 diabetes

Zhang

Zhao

et al. 2022

Endocrine Connections

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Objective: The beneficial effect of angiotensin(1–7), via the activation of its receptor, MAS-1, has been noted in diabetes treatment; however, how angiotensin(1–7) or MAS-1 affects insulin secretion remains elusive and whether endogenous level of angiotensin(1–7) or MAS-1 is altered in diabetic individuals remains unexplored. We recently identified an important role of CFTR, a cAMP-activated Cl- channel, in regulation of insulin secretion. Here, we tested possible involvement of CFTR in mediating angiotensin(1–7)’s effect on insulin secretion and measured the level of angiotensin(1–7), MAS-1 as well as CFTR in the blood of individuals with or without type 2 diabetes. Methods: Angiotensin(1–7)/MAS-1/CFTR pathway was determined by specific inhibitors, gene manipulation, western blotting as well as insulin ELISA in a pancreatic β cell line, RINm5F. Human blood samples were collected from 333 individuals with (n=197) and without (n=136) type 2 diabetes. Angiotensin(1–7), MAS-1 and CFTR level in the human blood were determined by ELISA. Results: In RINm5F cells, angiotensin(1–7) induced intracellular cAMP increase, CREB activation, enhanced CFTR expression and potentiated glucose-stimulated insulin secretion, which were abolished by a selective CFTR inhibitor, RNAi-knockdown of CFTR, or inhibition of MAS-1. In human subjects, the blood levels of MAS-1 and CFTR, but not angiotensin(1–7), were significantly higher in individuals with type 2 diabetes as compared to those in non-diabetic healthy subjects. In addition, blood levels of MAS-1 and CFTR were in significant positive correlation in type-2-diabetic but not non-diabetic subjects. Conclusion: These results suggested MAS-1 and CFTR as key players in mediating angiotensin(1–7)-promoted insulin secretion in pancreatic β cells; MAS-1 and CFTR are positively correlated and both upregulated in type 2 diabetes.

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A Survey of Membrane Proteins in Human Serum

Cited by 4 publications

References 27 publications

The Role of GPR120 Receptor in Essential Fatty Acids Metabolism in Schizophrenia

The Role of GPR120 Receptor in Essential Fatty Acids Metabolism in Schizophrenia

A Routine ‘Top-Down’ Approach to Analysis of the Human Serum Proteome

Angiotensin(1–7) activates MAS-1 and upregulates CFTR to promote insulin secretion in pancreatic β-cells: the association with type 2 diabetes

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