Inverse labeling–mass spectrometry for the rapid identification of differentially expressed protein markers/targets

Introduction: The aim of this study was to investigate the modified proteins in methylene blue/light-treated frozen plasma (MB-FP) compared with fresh frozen plasma (FFP) in order to gain a better application of MB/light-treated plasma in clinic transfusion. Methods: MB-FP and FFP were collected from Changchun central blood station, and a trichloroacetic acid/acetone precipitation method was used to remove albumin for the enrichment of lower abundance proteins. The plasma protein in MB-FP and FFP were separated using two-dimensional gel electrophoresis (2-DE) and the differentially expressed protein spots were analyzed using mass spectrometry. Finally, the differentially expressed proteins were tested using Western blot and enzyme-linked immunosorbent assay (ELISA). Results: Approximately 14 differentially expressed protein spots were detected in the MB-FP, and FFP was chosen as the control. After 2-DE comparison analysis and mass spectrometry, 8 significantly differentially expressed protein spots were identified, corresponding to 6 different proteins, including complement C1r subcomponent (C1R), inter-alpha-trypsin inhibitor heavy chain H4 (ITI-H4), keratin, type II cytoskeletal 1 (KRT1), hemopexin (HPX), fibrinogen gamma chain (FGG), and transthyretin (TTR). Western blot showed no significant difference in the expression level of KRT1 between MB-FP and FFP (p > 0.05). Both Western blot and ELISA indicated that the level of HPX was significantly higher in FFP than in MB-FP (p < 0.05). Conclusion: This comparative proteomics study revealed that some significantly modified proteins occur in MB-FP, such as C1R, ITI-H4, KRT1, HPX, FGG, and TTR. Our findings provide more theoretical data for using MB-FP in transfusion medicine. However, the relevance of the data for the transfusion of methylene blue/light-treated plasma remains unclear. The exact modification of these proteins and the effects of these modified proteins on their functions and their effects in clinical plasma infusion need to be further studied.

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Differentially expressed protein markers in human submandibular and sublingual secretions

International Journal of Oncology ◽

10.3892/ijo.25.5.1423 ◽

2004 ◽

Cited By ~ 5

Author(s):

Shen Hu ◽

Patricia Denny ◽

Paul Denny ◽

Yongming Xie ◽

Joseph Loo ◽

...

Keyword(s):

Differentially Expressed ◽

Protein Markers ◽

Differentially Expressed Protein

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Application of proteomics for the identification of differentially expressed protein markers for Down syndrome in maternal plasma

Prenatal Diagnosis ◽

10.1002/pd.2040 ◽

2008 ◽

Vol 28 (8) ◽

pp. 691-698 ◽

Cited By ~ 45

Author(s):

Aggeliki Kolialexi ◽

George Th. Tsangaris ◽

Nikos Papantoniou ◽

Athanasios K. Anagnostopoulos ◽

Kostantinos Vougas ◽

...

Keyword(s):

Down Syndrome ◽

Maternal Plasma ◽

Differentially Expressed ◽

Protein Markers ◽

Differentially Expressed Protein

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Inverse15N-metabolic labeling/mass spectrometry for comparative proteomics and rapid identification of protein markers/targets

Rapid Communications in Mass Spectrometry ◽

10.1002/rcm.725 ◽

2002 ◽

Vol 16 (14) ◽

pp. 1389-1397 ◽

Cited By ~ 30

Author(s):

Y. Karen Wang ◽

Zhixiang Ma ◽

Douglas F. Quinn ◽

Emil W. Fu

Keyword(s):

Mass Spectrometry ◽

Rapid Identification ◽

Comparative Proteomics ◽

Metabolic Labeling ◽

Protein Markers

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VDAC1 and RhoA in Plasma Exosomes Influence the Severity of Adolescent Idiopathic Scoliosis

10.21203/rs.3.rs-65485/v1 ◽

2020 ◽

Author(s):

Huizhen Li ◽

Nan Shen ◽

Lin Mao ◽

Meijia Chen ◽

Xuan Zhou ◽

...

Keyword(s):

Mass Spectrometry ◽

Liquid Chromatography ◽

Adolescent Idiopathic Scoliosis ◽

Tandem Mass Spectrometry ◽

Idiopathic Scoliosis ◽

Differentially Expressed ◽

Tandem Mass ◽

Healthy Controls ◽

Differentially Expressed Proteins ◽

Differentially Expressed Protein

Abstract Background: Adolescent idiopathic scoliosis (AIS) is the most common spine deformity, but biomarkers for its condition are lacking. Rhodopsin A (RhoA) and voltage-dependent anion-selective channel 1 (VDAC1) in plasma exosomes were defined as differentially expressed proteins between AIS patients and healthy controls. The purpose of this study was to assess exosomes as biomarkers for the occurrence and progression of AIS. Methods：We recruited 10 AIS patients and 8 healthy controls to detect expressed proteins from plasma by liquid chromatography coupled to tandem mass spectrometry. Plasma samples were analyzed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Pathway analysis identified that the VDAC1 and RhoA proteins were alterations expressed in the AIS patients, with the most different alteration was found in extracellular exosomes. Ultracentrifugation was carried out to isolate exosomes from plasma. Verification of the most differentially expressed protein was accessed by Western blot analysis and bioinformatics analysis was performed to predict the pathway of it.Results: 42 of significantly differentially expressed proteins were found in all subjects, and 17 proteins had significant difference. The differentially expressed proteins were enriched in plasma exosomes, and some proteins, such as FN1, were upregulated and others, such as VDAC1, RhoA and AHNAK, were downregulated in the AIS patients. Furthermore, ultracentrifugation was carried out to isolate exosomes from plasma, and RhoA and VDAC1 proteins in plasma exosomes were verified to downregulate by western blot. KEGG signaling pathways were used to predict potential pathways involved in the RhoA and VDAC1 proteins in the AIS patients. We found that the RhoA protein influences AIS probably through the chemokine signaling pathway, platelet activation and cAMP signaling pathway, and the VDAC1 protein is a key factor that participates in the necroptosis pathway, acting on the development of AIS.Conclusions: Consequently, this study mapped a profile of plasma protein, found the differentially expressed protein in AIS, which indicating that plasma exosomes, as a novel biomarker with high specificity, could be associated with the severity of AIS.

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