Proteomic Analysis of a Membrane Skeleton Fraction from Human Liver

He, Jintang; Liu, Yashu; He, Sizhi; Wang, Qingsong; Hai, Pham Dang; Ji, Jianguo

doi:10.1021/pr070197v

Cited by 20 publications

(28 citation statements)

References 56 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Our results suggest that the relationship between membrane disruption efficiency (Table 2) and the physicochemical properties of ASB detergents (CMC, HLB) can explain the improvement in the solubilization of membrane proteins, as described in proteomic research papers (11)(12)(13)(14)(15)(16)22,23).…”

Section: Hemolytic Experimentssupporting

confidence: 63%

“…The zwitterionic amidosulfobetaine detergents ASB-14 and ASB-16 (linear alkylamido propyl dimethyl amino propanosulfonates) with 14 and 16 atoms of carbon in the acyl chain, respectively, were synthesized by Chevallet et al (11). They have contributed to the advance of two-dimensional gel electrophoresis analysis, improving the solubilization of many hydrophobic proteins such Erythrocyte solubilization by ASB detergents www.bjournal.com.br as erythrocyte band III (12,13) and others (11,(14)(15)(16).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Solubilization of human erythrocyte membranes by ASB detergents

Domingues

Malheiros

Paula

2008

Braz J Med Biol Res

View full text Add to dashboard Cite

Understanding the membrane solubilization process and finding effective solubilizing agents are crucial challenges in biochemical research. Here we report results on the interaction of the novel linear alkylamido propyl dimethyl amino propanosulfonate detergents, ASB-14 and ASB-16, with human erythrocyte membranes. An estimation of the critical micelle concentration of these zwitterionic detergents (ASB-14 = 100 µM and ASB-16 = 10 µM) was obtained using electron paramagnetic resonance. The amount of proteins and cholesterol solubilized from erythrocytes by these detergents was then determined. The hemolytic activities of the ASB detergents were assayed and the detergent/lipid molar ratios for the onset of hemolysis (R e sat ) and total lysis (R e sol ) were calculated, allowing the determination of the membrane binding constants (K b ). ASB-14 presented lower membrane affinity (K b = 7050 M -1 ) than ASB-16 (K b = 15610 M -1 ). The amount of proteins and cholesterol solubilized by both ASB detergents was higher while R e sat values (0.22 and 0.08 detergent/lipid for ASB-14 and ASB-16, respectively) were smaller than those observed with the classic detergents CHAPS and Triton X-100. These results reveal that, besides their well-known use as membrane protein solubilizers to enhance the resolution of two dimensional electrophoresis/ mass spectrometry, ASB-14 and ASB-16 are strong hemolytic agents. We propose that the physicochemical properties of ASB detergents determine their membrane disruption efficiency and can help to explain the improvement in the solubilization of membrane proteins, as reported in the literature.

show abstract

Section: Hemolytic Experimentssupporting

confidence: 63%

Section: Introductionmentioning

confidence: 99%

Solubilization of human erythrocyte membranes by ASB detergents

Domingues

Malheiros

Paula

2008

Braz J Med Biol Res

View full text Add to dashboard Cite

show abstract

“…Western Blotting-Western blotting was performed essentially as described before (32). Briefly, 16 g of proteins from the undifferentiated and differentiated HSR-GBM1 cells were separated by 4 -20% SDS-PAGE and transferred to polyvinylidene difluoride membranes (Bio-Rad, CA).…”

Section: Methodsmentioning

confidence: 99%

CD90 is Identified as a Candidate Marker for Cancer Stem Cells in Primary High-Grade Gliomas Using Tissue Microarrays

Liu

Zhu

et al. 2012

Molecular & Cellular Proteomics

Self Cite

125

108

View full text Add to dashboard Cite

Although CD90 has been identified as a marker for various kinds of stem cells including liver cancer stem cells (CSCs) that are responsible for tumorigenesis, the potential role of CD90 as a marker for CSCs in gliomas has not been characterized. To address the issue, we investigated the expression of CD90 in tissue microarrays containing 15 glioblastoma multiformes (GBMs), 19 WHO grade III astrocytomas, 13 WHO grade II astrocytomas, 3 WHO grade I astrocytomas and 8 normal brain tissues. Immunohistochemical analysis showed that CD90 was expressed at a medium to high level in all tested high-grade gliomas (grade III and GBM) whereas it was barely detectable in low-grade gliomas (grade I and grade II) and normal brains. Double immunofluorescence staining for CD90 and CD133 in GBM tissues revealed that CD133 ؉ CSCs are a subpopulation of CD90؉ cells in GBMs in vivo. Flow cytometry analysis of the expression of CD90 and CD133 in GBM-derived stem-like neurospheres further confirmed the conclusion in vitro. The expression levels of both CD90 and CD133 were reduced along with the loss of stem cells after differentiation. Furthermore, the limiting dilution assay demonstrated that the sphere formation ability was comparable between the CD90 ؉ /CD133 ؉ and the CD90 ؉ /CD133 ؊ populations of GBM neurospheres, which is much higher than that of the CD90 ؊ /CD133 ؊ population. We also performed double staining for CD90 and a vascular endothelial cell marker CD31 in tissue microarrays which revealed that the CD90 ؉ cells were clustered around the tumor vasculatures in high-grade glioma tissues. These findings suggest that CD90 is not only a potential prognostic marker for high-grade gliomas but also a marker for CSCs within gliomas, and it resides

show abstract

“…Western Blotting-Western blotting was performed essentially as described previously (18). Antibodies used in this study include: antiornithine aminotransferase generously provided by Dr. Olivier Levillain (Universite Claude Bernard Lyon 1) (19), anti-Oxphos complex I 39-kDa subunit from Molecular Probes (Eugene, OR), and anti-glutamine synthetase and anti-peroxisome proliferator-activated receptor ␣ (PPAR␣) from Abcam (Cambridge, UK).…”

Section: Methodsmentioning

confidence: 99%

Comparative Studies of Early Liver Dysfunction in Senescence-accelerated Mouse Using Mitochondrial Proteomics Approaches

Liu

et al. 2008

Molecular & Cellular Proteomics

Self Cite

View full text Add to dashboard Cite

The liver is a complex and unique organ responsible for a breadth of functions crucial to sustaining life, especially for various metabolic processes in its mitochondria. Senescence-accelerated mouse prone/8 (SAMP8), a widely used aging model, exhibits an oxidative stress-induced aging phenotype and severe mitochondria-related liver pathology that are not seen in senescence-accelerated mouse resistant/1 (SAMR1). Here we used both two-dimensional electrophoresis-and ICAT-based mitochondrial proteomics analysis to view the liver mitochondrial protein alterations between SAMP8 and SAMR1. Compared with SAMR1, decreased expression and activity of mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase were detected in SAMP8 at 6 months old (SAMP8-6m). As the key enzyme of ketogenesis, 3-hydroxy-3-methylglutaryl-CoA synthase is well known to be transcriptionally regulated by peroxisome proliferator-activated receptor ␣, which was also expressed at lower levels in SAMP8-6m livers. In addition, down-regulation of two peroxisome proliferator-activated receptor ␣ target gene products (acyl-CoA oxidase and enoyl-CoA hydratase), elevation of triglyceride, and reduction of acetyl-CoA were observed, indicating abnormal fatty acid metabolism in SAMP8-6m livers. In addition eight proteins (NDUAA, NDUBA, NDUB7, NDUS1, NDUS3, NDUV1, ETFA, and UCRI) of mitochondrial complexes were down-regulated in SAMP8-6m, resulting in mitochondria-related liver dysfunction characterized by enhanced oxidative stress-induced molecular damage (lipid peroxide and oxidized protein) and depressed energy production (ATP). Glutamine synthetase and ornithine aminotransferase involved in glutamine synthesis were up-regulated in SAMP8 livers at both 1 and 6 months old that may be related to the accumulation of glutamate and glutamine. Our work provided useful clues to understanding the molecular mechanism underlying liver dysfunction in senescence-accelerated mouse. Molecular & Cellular Proteomics 7:1737-1747, 2008.The liver is regarded as the major organ for a number of physiological presses. As the main energy producers, liver mitochondria are considered to be the central integrators of intermediary metabolism including fatty acid oxidation, the Krebs cycle, oxidative phosphorylation, ketogenesis, and the urea cycle. Furthermore most of the liver disorders have been reported to have increased reactive oxygen species (ROS) 1 and decreased ATP as vital characteristics (1, 2), which are both generated through the mitochondrial respiratory chain.The senescence-accelerated mouse (SAM) model is an aging model obtained by continuous sister-brother breeding from original litters of AKR/J mice (3). SAM includes two strains, i.e. SAM prone (SAMP) and SAM resistant (SAMR). SAMP exhibit a shortened life span and early manifestation of various symptoms of senescence, whereas SAMR are senescence-resistant inbred strains (4). SAMP8, a substrain of SAMP, has become a major biogerontological resource in aging research with SAMR1 as control (5, 6).Recently Ye et al. (7) h...

show abstract

Proteomic Analysis of a Membrane Skeleton Fraction from Human Liver

Cited by 20 publications

References 56 publications

Solubilization of human erythrocyte membranes by ASB detergents

Solubilization of human erythrocyte membranes by ASB detergents

CD90 is Identified as a Candidate Marker for Cancer Stem Cells in Primary High-Grade Gliomas Using Tissue Microarrays

Comparative Studies of Early Liver Dysfunction in Senescence-accelerated Mouse Using Mitochondrial Proteomics Approaches

Contact Info

Product

Resources

About