Generation of novel Id2 and E2-2, E2A and HEB antibodies reveals novel Id2 binding partners and species-specific expression of E-proteins in NK cells
| Autor: | Nicholas D. Huntington, Jai Rautela, Lynn M. Corcoran, Rebecca B. Delconte, Andrew I. Webb, Joanna R Groom, Ngaire Elwood, Angaleena Anthony, Wilford Goh, Elliot Surgenor, Tobias Kratina, Fernando Souza-Fonseca-Guimaraes, Laura F. Dagley |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Immunoprecipitation Immunology Immunofluorescence Antibodies Cell Line Mice 03 medical and health sciences Transcription Factor 4 0302 clinical medicine Basic Helix-Loop-Helix Transcription Factors medicine Animals Humans Cytotoxic T cell Cell Lineage Molecular Biology Transcription factor Inhibitor of Differentiation Protein 2 biology medicine.diagnostic_test Chromatin Cell biology Killer Cells Natural Mice Inbred C57BL 030104 developmental biology Interleukin 15 Cell culture biology.protein Antibody 030215 immunology |
| Zdroj: | Molecular Immunology. 115:56-63 |
| ISSN: | 0161-5890 |
| Popis: | NK cells are cytotoxic lymphocytes with a key role in limiting tumour metastases. In mice, the NK cell lineage continually expresses high levels of the Inhibitor of DNA-binding 2 (Id2) protein and loss of Id2 is incongruous with their survival due to aberrant E-protein target gene activity. Using novel Id2 and E-protein antibodies that detect both mouse and human proteins, we have extensively characterised Id2 and E-protein expression in murine and human NK cells. We detected clear expression of E2 A and HEB, and to a lesser extent E2-2 in murine NK cells. In contrast HEB appears to be the major E-protein expressed in human NK cells, with minor E2-2 expression and surprisingly, no E2 A detected in primary NK cells nor human NK cell lines. These novel antibodies are also functional in immunofluorescence and immunoprecipitation. Mass spectrometry analysis of Id2 immuno-precipitated from murine NK cells revealed a number of novel associated proteins including several members of the SWI/SNF-related matrix-associated actin-dependent regulator chromatin (SMARC) and Mediator complex (MED) families. Taken together, these data highlight the utility of novel Id2 and E-protein antibodies and caution against mouse models for understanding Id2/E-protein biology in NK cells given their clearly disparate expression patternbetween species. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect