62,784 research outputs found
Hybridoma cells containing intracellular anti-ricin antibodies show ricin meets secretory antibody before entering the cytosol
No full textHybridoma cells which synthesize monoclonal antibodies (mAb) that block ricin toxicity were 50-300-fold resistant to ricin compared with other hybridomas. Two of the mAb blocked two isozymes of ricin, D and E, to different and opposite extents, and the hybridoma cell resistance to the two forms of ricin closely corresponded with the mAb reactivity. The hybridoma cell resistance to ricin was therefore due to the binding activity of the mAb produced by the cells. Neither rabbit polyclonal antibodies, which neutralized extracellular anti-ricin mAb, nor quantitative removal of hybridoma cell surface IgG with papain affected the cellular resistance to ricin. Therefore, neither extracellular or cell surface antibodies contributed to the resistance of the hybridoma cells. In contrast, inhibition of protein synthesis by cycloheximide or puromycin, which selectively decreased levels of intracellular secretory IgG, decreased the hybridoma cell resistance to ricin. We conclude that intracellular mAb, synthesized de novo for subsequent secretion, block ricin toxicity. Ricin therefore must meet intracellular secretory antibodies before reaching the cytosol. The monoclonal antibodies can also be used to study toxin function within intracellular compartments. An antibody specific for the galactose-binding site of ricin blocks ricin intracellularly, showing that the ricin galactose-binding activity is required in an intracellular compartment for transport of ricin A chain to the cytosol
Cloned fragment of diphtheria toxin linked to T cell-specific antibody identifies regions of B chain active in cell entry
No full textThe role of discrete domains of diphtheria toxin (DT) B chain in cytosol entry and cytotoxicity was investigated by linking a monoclonal antibody recognizing the human T cell-specific antigen T3 (UCHT1) to diphtheria toxin (UCHT1-DT), DT A subunit (UCHT1-DTA), or to a genetically engineered form of DT (UCHT1-MspSA) lacking the C-terminal 17-kDa portion of the B subunit. The N-terminal 21-kDa region of DT B chain increased toxicity of UCHT1-DTA 100-fold (UCHT1-MspSA) while addition of the C-terminal 17-kDa region (UCHT1-DT) increased toxicity 100-fold more. The cytotoxicity was dependent upon antibody binding as demonstrated by blocking toxicity with excess UCHT1. The differences in toxicity between these reagents were not due to differences in ADP-ribosylation activity of DT A chain, binding activity of the antibody moiety, extent of DT nicking, or the cross-linking method, so we conclude that the large differences in toxicity were due to the presence of different B chain domains. The large increase in toxicity by the C-terminal region of DT B did not appear to be caused by DT receptor binding. The lysosomotropic agent NH4Cl blocked the cytotoxic effect of DT, UCHT1-DT, and UCHT1-MspSA but not UCHT1-DTA
Letter from J. R. Eakin to Arthur G. Ringland
No full textLetter (copy) from J. R. Eakin to Arthur C. Ringland about the alignment of 40 acres near the Buggeln ranch
Letter from Arno B. Cammerer to J. R. Eakin
No full textLetter from Arno B. Cammerer to J. R. Eakin describing the procedure for purchasing Bright Angel Trail
Letter from J. R. Eakin to Carl Hayden
No full textLetter from J. R. Eakin to Carl T. Hayden concerning access to Rowe Well and the canyon
Letter from J. R. Eakin to Stephen Mather
No full textLetter from J. R. Eakin to Stephen T. Mather about expenses and reconstruction of the Kaibab Trail
Mitochondrial release of AIF and EndoG requires caspase activation downstream of Bax/Bak-mediated permeabilization
No full textMitochondrial outer-membrane permeabilization by pro-apoptotic Bcl-2 family members plays a crucial role in apoptosis induction. However, whether this directly causes the release of the different mitochondrial apoptogenic factors simultaneously is currently unknown. Here we report that in cells or with isolated mitochondria, pro-apoptotic Bcl-2 proteins cause the release of cytochrome c, Smac/Diablo and HtrA2/Omi but not endonuclease G (EndoG) and apoptosis-inducing factor (AIF). In cells treated with Bax/Bak-dependent pro-apoptotic drugs, neither the caspase inhibitor zVAD-fmk nor loss of Apaf-1 affected the efflux of cytochrome c, Smac/Diablo and HtrA2/Omi, but both prevented the release of EndoG and AIF. Our findings identify the mitochondrial response to pro-apoptotic stimuli as a selective process leading to a hierarchical ordering of the effectors involved in cell death induction. Moreover, as in Caenorhabditis elegans, EndoG and AIF act downstream of caspase activation. Thus EndoG and AIF seem to define a 'caspase-dependent' mitochondria-initiated apoptotic DNA degradation pathway that is conserved between mammals and nematodes
Letter from Carl Hayden to J. R. Eakin
No full textLetter from Carl T. Hayden to J. R. Eakin regarding changes to the Grand Canyon National Park boundaries and the purchase of lands from William Randolph Hearst
Letter from J. R. Eakin to Stephen Mather
No full textLetter from J. R. Eaking to the National Park Service director about changes to the Grand Canyon National Park boundaries, and access to water near the Buggeln property on Desert View road
[Letter from J. R. Roberts to Sister, November 24, 1878]
No full textLetter from J. R. Roberts to sister. J. R. thanked his sister for gifts that were sent and went on to update her on what was happening in their families' lives. The letter ended with a mention that people were searching for land claims in the area and the author wanted their mother to not worry about them
- …
