mouse cd31 pecam 1 protein  (MedChemExpress)

Journal: Journal of Biological Chemistry

Article Title: Interaction of Platelet Endothelial Cell Adhesion Molecule (PECAM) with α2,6-Sialylated Glycan Regulates Its Cell Surface Residency and Anti-apoptotic Role

doi: 10.1074/jbc.m114.563585

Figure Lengend Snippet: FIGURE 1. Sialylated oligosaccharides inhibit the homophilic mouse PECAM interaction. A, PECAM-His and PECAM-Fc pulled down by Dynabeads protein G in the presence of a series of oligosaccharides (2-(trimethylsilyl)ethyl (SE) glycoside forms, 0, 0.2, 1, and 5 mM) were evaluated by Western blot analysis. WB, Western blot. B, the relative levels of PECAM-His bound to immobilized PECAM-Fc in the presence of a series of oligosaccharides (0.2, 1, and 5 mM) were quantified by Western blot analysis. The data are shown as the means S.E. when the level of PECAM-His in the absence of an oligosaccharide was set at 100% (n 3).

Article Snippet: Materials—The sources of the materials used in this study were as follows: tissue culture media and reagents, including DMEM and Dynabeads protein G from Invitrogen, Arthrobacter ureafaciens sialidase from Nacalai Tesque, Vibrio cholera sialidase and recombinant peptide N-glycosidase F from New England Biolabs, recombinant mouse PECAM from R&D Systems, human PECAM-His from Reprokine, nickel-Sepharose and Sephadex G-10 from GE Healthcare, protein molecular weight standards from Bio-Rad, and all other chemicals from Sigma or Wako Chemicals.

Techniques: Western Blot

Journal: Journal of Biological Chemistry

Article Title: Interaction of Platelet Endothelial Cell Adhesion Molecule (PECAM) with α2,6-Sialylated Glycan Regulates Its Cell Surface Residency and Anti-apoptotic Role

doi: 10.1074/jbc.m114.563585

Figure Lengend Snippet: FIGURE 2. 2,6-Sialylated glycocluster probe binds to immobilized PECAM and endothelial cells. A, schematic structure of the sialylated glyco- cluster probes used in this study. The 2,3-type disialylated, 2,6-type disia- lylated, or asialo-type N-glycan clusters were labeled with nitrobenzoxadia- zole (star) (30). B, human asialo-PECAM-His immobilized to nickel-agarose beads was incubated with the three glycocluster probes. After a brief wash the beads were observed by fluorescence microscopy. Scale bar, 200 m. C, prefixed HUVECs were incubated with the series of glycocluster probes and observedbyfluorescencemicroscopy.Arrowheadsindicatecellborders.Scale bar, 20 m.

Article Snippet: Materials—The sources of the materials used in this study were as follows: tissue culture media and reagents, including DMEM and Dynabeads protein G from Invitrogen, Arthrobacter ureafaciens sialidase from Nacalai Tesque, Vibrio cholera sialidase and recombinant peptide N-glycosidase F from New England Biolabs, recombinant mouse PECAM from R&D Systems, human PECAM-His from Reprokine, nickel-Sepharose and Sephadex G-10 from GE Healthcare, protein molecular weight standards from Bio-Rad, and all other chemicals from Sigma or Wako Chemicals.

Techniques: Glycoproteomics, Labeling, Incubation, Fluorescence, Microscopy

Journal: Journal of Biological Chemistry

Article Title: Interaction of Platelet Endothelial Cell Adhesion Molecule (PECAM) with α2,6-Sialylated Glycan Regulates Its Cell Surface Residency and Anti-apoptotic Role

doi: 10.1074/jbc.m114.563585

Figure Lengend Snippet: FIGURE 3. Mouse PECAM mainly possesses sialylated biantennary N-gly- cans. A, a series of mutant Fc-PECAMs in which each potential N-glycosyla- tion site was mutated were purified from overexpressing COS cells, subjected to SDS-PAGE, and visualized by silver-staining. B, immunopurified PECAM (arrowhead) from mouse lung tissues was verified by SDS-PAGE and silver staining. The de-N-glycosylated anti-PECAM IgG used for immunoaffinity chromatography is also shown (right lane). The only contaminant except for IgG, shown by the asterisk, was shown not to have any N-glycans because it was resistant to peptide N-glycosidase F treatment. C, PA-labeled N-glycans releasedfromPECAMwereseparatedbyanion-exchangeHPLC.Thenumbers shown indicate the eluted positions of standard N-glycans having 0, 1, 2, 3, and 4 sialic acid residues, respectively.

Article Snippet: Materials—The sources of the materials used in this study were as follows: tissue culture media and reagents, including DMEM and Dynabeads protein G from Invitrogen, Arthrobacter ureafaciens sialidase from Nacalai Tesque, Vibrio cholera sialidase and recombinant peptide N-glycosidase F from New England Biolabs, recombinant mouse PECAM from R&D Systems, human PECAM-His from Reprokine, nickel-Sepharose and Sephadex G-10 from GE Healthcare, protein molecular weight standards from Bio-Rad, and all other chemicals from Sigma or Wako Chemicals.

Techniques: Mutagenesis, Purification, SDS Page, Silver Staining, Chromatography, Labeling

Journal: Journal of Biological Chemistry

Article Title: Interaction of Platelet Endothelial Cell Adhesion Molecule (PECAM) with α2,6-Sialylated Glycan Regulates Its Cell Surface Residency and Anti-apoptotic Role

doi: 10.1074/jbc.m114.563585

Figure Lengend Snippet: FIGURE 4. Disialylated N-glycan is found in PECAM purified from mouse lungs. A–C, MS2 spectra at m/z 1241.0 [M2H]2 (A), m/z 1167.9 [M2H]2 (B), and m/z 1263.5 [MHNa]2 (C) are shown in the positive ion mode.

Article Snippet: Materials—The sources of the materials used in this study were as follows: tissue culture media and reagents, including DMEM and Dynabeads protein G from Invitrogen, Arthrobacter ureafaciens sialidase from Nacalai Tesque, Vibrio cholera sialidase and recombinant peptide N-glycosidase F from New England Biolabs, recombinant mouse PECAM from R&D Systems, human PECAM-His from Reprokine, nickel-Sepharose and Sephadex G-10 from GE Healthcare, protein molecular weight standards from Bio-Rad, and all other chemicals from Sigma or Wako Chemicals.

Techniques: Glycoproteomics, Purification

Journal: Journal of Biological Chemistry

Article Title: Interaction of Platelet Endothelial Cell Adhesion Molecule (PECAM) with α2,6-Sialylated Glycan Regulates Its Cell Surface Residency and Anti-apoptotic Role

doi: 10.1074/jbc.m114.563585

Figure Lengend Snippet: FIGURE 5. Sialidase or 2,6-sialylated oligosaccharide treatment causes PECAM internalization in endothelial cells. A, HUVECs were treated with V. cholera sialidase, fixed, and stained with anti-early endosome antigen 1 (EEA1; green) and PECAM (red) antibodies and DAPI (blue). B, HUVECs were incubated with 2 mM lactose, 2,3- or 2,6-sialylated lactose, or 2,6-sialy- lated or asialo-biantennary N-glycan for 18 h, fixed, and stained with PECAM (red) antibody. Scale bar, 20 m.

Article Snippet: Materials—The sources of the materials used in this study were as follows: tissue culture media and reagents, including DMEM and Dynabeads protein G from Invitrogen, Arthrobacter ureafaciens sialidase from Nacalai Tesque, Vibrio cholera sialidase and recombinant peptide N-glycosidase F from New England Biolabs, recombinant mouse PECAM from R&D Systems, human PECAM-His from Reprokine, nickel-Sepharose and Sephadex G-10 from GE Healthcare, protein molecular weight standards from Bio-Rad, and all other chemicals from Sigma or Wako Chemicals.

Techniques: Staining, Incubation, Glycoproteomics

Journal: Journal of Biological Chemistry

Article Title: Interaction of Platelet Endothelial Cell Adhesion Molecule (PECAM) with α2,6-Sialylated Glycan Regulates Its Cell Surface Residency and Anti-apoptotic Role

doi: 10.1074/jbc.m114.563585

Figure Lengend Snippet: FIGURE7.PECAMisalectinthatpreferssialylatedglycans.Thelectinprop- erty of PECAM toward sialylated glycans could be important not only for its homophilic interaction but also for interactions with other endothelial sur- face molecules. The N-terminal Ig domain mediates the homophilic PECAM interaction, whereas multiple Ig domains are involved in the heterophilic PECAMinteraction(17).WhichIgdomainisresponsibleforthelectinproperty of PECAM remains to be studied. ITIM, immunoreceptor tyrosine inhibitory motif.

Article Snippet: Materials—The sources of the materials used in this study were as follows: tissue culture media and reagents, including DMEM and Dynabeads protein G from Invitrogen, Arthrobacter ureafaciens sialidase from Nacalai Tesque, Vibrio cholera sialidase and recombinant peptide N-glycosidase F from New England Biolabs, recombinant mouse PECAM from R&D Systems, human PECAM-His from Reprokine, nickel-Sepharose and Sephadex G-10 from GE Healthcare, protein molecular weight standards from Bio-Rad, and all other chemicals from Sigma or Wako Chemicals.

Techniques: