HAM: A new epitope-tag for in vivo protein labeling

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

URL: http://www.ncbi.nlm.nih.gov/pubmed/11455955
DOI: 10.1023/A:1011008018565

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: http://repository.cshl.edu/id/eprint/29395

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