Herbst, A., Tansey, W. P. (2000) HAM: A new epitope-tag for in vivo protein labeling. Molecular Biology Reports, 27 (4). pp. 203-208. ISSN 0301-4851
Abstract
The ability to metabolically label proteins with S-35-methionine is critical for the analysis of protein synthesis and turnover. Despite the importance of this approach, however, efficient labeling of proteins in vivo is often limited by a low number of available methionine residues, or by deleterious side-effects associated with protein overexpression. To overcome these limitations, we have created a methionine-rich variant of the widely used HA tag, called HAM, for use with ectopically expressed proteins. Here we describe the development of a series of vectors, and corresponding antisera, for the expression and detection of HAM-tagged proteins in mammalian cells. We show that the HAM tag dramatically improves the sensitivity of S-35-methionine labeling, and permits the analysis of Myc oncoprotein turnover even when HAM-tagged Myc is expressed at levels comparable to that of the endogenous protein. Because of the improved sensitivity provided by the HAM tag, the vectors and antisera described here should be useful for the analysis of protein synthesis and destruction at physiological levels of protein expression.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | antibody epitope-tag metabolic labeling protein stability ACTIVATION DOMAINS CELL PROTEOLYSIS MYC GROWTH OCT-1 |
| Subjects: | Investigative techniques and equipment bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification |
| CSHL Authors: | |
| Communities: | CSHL labs > Tansey lab |
| Depositing User: | Matt Covey |
| Date: | 2000 |
| Date Deposited: | 30 Jan 2014 22:17 |
| Last Modified: | 30 Jan 2014 22:17 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/29395 |
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