Identification of aquaporin-5 and lipid rafts in human resting saliva and their release into cevimeline-stimulated saliva

Pan, Yan; Iwata, Fusako; Wang, Di; Muraguchi, Masahiro; Ooga, Keiko; Ohmoto, Yasukazu; Takai, Masaaki; Cho, Gota; Kang, Jin-sen; Shono, Masayuki; Li, Xuejun; Okamura, Ko; Mori, Toyoki; Ishikawa, Yasuko

doi:10.1016/j.bbagen.2008.08.009

Cited by 26 publications

(21 citation statements)

References 35 publications

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“…In addition, the Brij 98 treatment dramatically increased the amount of AQP0 and AQP5 in the low-density fraction. Increasing evidence supports the lipid raft localization of AQP5, [29][30][31] whereas the localization of AQP0 in lipid rafts has been reported previously. 16,17 These results suggest that Brij 98 provides better yields of lipid raft marker proteins in the low-density fractions; therefore, further studies were performed only on DRM isolated after Brij 98 solubilization.…”

Section: Isolation Of Lipid Raft-like Drmsmentioning

confidence: 92%

Proteomic Analysis of Lipid Raft-Like Detergent-Resistant Membranes of Lens Fiber Cells

Wang

Schey

2015

Invest. Ophthalmol. Vis. Sci.

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Citation: Wang Z, Schey KL. Proteomic analysis of lipid raft-like detergentresistant membranes of lens fiber cells. Invest Ophthalmol Vis Sci. 2015;56:8349-8360. DOI:10.1167/ iovs.15-18273 PURPOSE. Plasma membranes of lens fiber cells have high levels of long-chain saturated fatty acids, cholesterol, and sphingolipids-key components of lipid rafts. Thus, lipid rafts are expected to constitute a significant portion of fiber cell membranes and play important roles in lens biology. The purpose of this study was to characterize the lens lipid raft proteome.METHODS. Quantitative proteomics, both label-free and iTRAQ methods, were used to characterize lens fiber cell lipid raft proteins. Detergent-resistant, lipid raft membrane (DRM) fractions were isolated by sucrose gradient centrifugation. To confirm protein localization to lipid rafts, protein sensitivity to cholesterol removal by methyl-b-cyclodextrin was quantified by iTRAQ analysis. RESULTS.A total of 506 proteins were identified in raft-like detergent-resistant membranes. Proteins identified support important functions of raft domains in fiber cells, including trafficking, signal transduction, and cytoskeletal organization. In cholesterol-sensitivity studies, 200 proteins were quantified and 71 proteins were strongly affected by cholesterol removal. Lipid raft markers flotillin-1 and flotillin-2 and a significant fraction of AQP0, MP20, and AQP5 were found in the DRM fraction and were highly sensitive to cholesterol removal. Connexins 46 and 50 were more abundant in nonraft fractions, but a small fraction of each was found in the DRM fraction and was strongly affected by cholesterol removal. Quantification of modified AQP0 confirmed that fatty acylation targeted this protein to membrane raft domains.CONCLUSIONS. These data represent the first comprehensive profile of the lipid raft proteome of lens fiber cells and provide information on membrane protein organization in these cells.

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Section: Isolation Of Lipid Raft-like Drmsmentioning

confidence: 92%

Proteomic Analysis of Lipid Raft-Like Detergent-Resistant Membranes of Lens Fiber Cells

Wang

Schey

2015

Invest. Ophthalmol. Vis. Sci.

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“…Route 3 is the trafficking route for AQP5 to the nucleus together with cholesterol-poor and sphingolipid-rich microdomains (shown in this study). In route 4, AQP5 is released into saliva by exosomes [12,13]. Route 5 is the trafficking route for AQP5 together with zymogen granules [59,60].…”

Section: Closedmentioning

confidence: 99%

“…The activation of M 3 -and M 1 -mAChRs induces the rapid translocation of AQP5, together with lipid rafts, from the apical cytoplasm to the APM by enhancement of [Ca 2+ ]i [10,11]. AQP5 and lipid rafts are then released into saliva [12]. AQP5 is known to be present in parotid gland exosomes [13,14], which accumulate within multivesicular bodies and are released into saliva from parotid cells upon fusion of multivesicular bodies with the plasma membrane in a Ca 2+ -triggered reaction [15,16].…”

Section: Introductionmentioning

confidence: 99%

Activation of muscarinic receptors in rat parotid acinar cells induces AQP5 trafficking to nuclei and apical plasma membrane

Cho

Bragiel

Wang

et al. 2015

Biochimica et Biophysica Acta (BBA) - General Subjects

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“…The membrane was then washed three times with TBS-T for 30 min and incubated at room temperature for 1 h with diluted (1:2000) secondary AP-labeled IgG (Santa Cruz Biotechnology, Santa Cruz, CA, USA). Blots were extensively washed in TBST buffer and detection was done using the NBT/BCIP reaction (Amersco, NJ, USA) [7] .…”

Section: Western Blot Analysismentioning

confidence: 99%

Roles of vimentin and 14-3-3 zeta/delta in the inhibitory effects of heparin on PC-3M cell proliferation and B16-F10-luc-G5 cells metastasis

et al. 2012

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Aim: To investigate the inhibitory effects of heparin on PC-3M cells proliferation in vitro and B16-F10-luc-G5 cells metastasis in Balb/c nude mice and identify the protein expression patterns to elucidate the action mechanism of heparin. Methods: Human prostate cancer PC-3M cells were incubated with heparin 0.5 to 125 μg/mL for 24 h. The proliferation of PC-3M cells was assessed by MTS assay. BrdU incoporation and Ki67 expression were detected using a high content screening (HCS) assay. The cell cycle and apoptosis of PC-3M cells were tested by flow cytometry. B16-F10-luc-G5 cardinoma cells were injected into the lateral tail vein of 6-week old male Balb/c nude mice and heparin 30 mg/kg was administered iv 30 min before and 24 h after injection. The metasis of B16-F10-luc-G5 cells was detected by bioluminescence assay. Activated partial thromboplastin time (APTT) and hemorheological parameters were measured on d 14 after injection of B16-F10-luc-G5 carcinoma cells in Balb/c mice. The global protein changes in PC-3M cells and frozen lung tissues from mice burdened with B16-F10-luc-G5 cells were determined by 2-dimensional gel electrophoresis and image analysis. The protein expression of vimentin and 14-3-3 zeta/delta was measured by Western blot. The mRNA transcription of vimentin, transforming growth factor (TGF)-β, E-cadherin, and α v -integrin was measured by RT-PCR. Results: Heparin 25 and 125 μg/mL significantly inhibited the proliferation, arrested the cells in G 1 phase, and suppressed BrdU incorporation and Ki67 expression in PC-3M cells compared with the model group. But it had no significant effect on apoptosis of PC-3M cells. Heparin 30 mg/kg markedly inhibits the metastasis of B16-F10-luc-G5 cells on day 8. Additionally, heparin administration maintained relatively normal red blood hematocrit but had no influence on APTT in nude mice burdened with B16-F10-luc-G5 cells. Thirty of down-regulated protein spots were identified after heparin treatment, many of which are related to tumor development, extracellular signaling, energy metabolism, and cellular proliferation. Vimentin and 14-3-3 zeta/delta were identified in common in PC-3M cells and the lungs of mice bearing B16-F10-luc-G5 carcinoma cells. Heparin 25 and 125 μg/mL decreased the protein expression of vimentin and 14-3-3 zeta/delta and the mRNA expression of α v -integrin. Heparin 125 μg/mL decreased vimentin and E-cadherin mRNA transcription while increased TGF-β mRNA transcription in the PC-3M cells, but the differences were not significant. Transfection of vimentin-targeted siRNA for 48 h significantly decreased the BrdU incoporation and Ki67 expression in PC-3M cells. Conclusion: Heparin inhibited PC-3M cell proliferation in vitro and B16-F10-luc-G5 cells metastasis in nude mice by inhibition of vimentin, 14-3-3 zeta/delta, and α v -integrin expression.

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Identification of aquaporin-5 and lipid rafts in human resting saliva and their release into cevimeline-stimulated saliva

Cited by 26 publications

References 35 publications

Proteomic Analysis of Lipid Raft-Like Detergent-Resistant Membranes of Lens Fiber Cells

Proteomic Analysis of Lipid Raft-Like Detergent-Resistant Membranes of Lens Fiber Cells

Activation of muscarinic receptors in rat parotid acinar cells induces AQP5 trafficking to nuclei and apical plasma membrane

Roles of vimentin and 14-3-3 zeta/delta in the inhibitory effects of heparin on PC-3M cell proliferation and B16-F10-luc-G5 cells metastasis

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