Journal article
Mass spectrometry identification of NASP binding partners in HeLa cells
Proteins, structure, function, and bioinformatics, Vol.61(1), pp.1-5
10/01/2005
DOI: 10.1002/prot.20600
PMID: 16080155
Abstract
Nuclear autoantigenic sperm protein (NASP) is a linker histone binding protein that is cell-cycle regulated. Synchronized HeLa cells are delayed in progression through the G1/S border when transiently transfected to overexpress full-length NASP, but not the histone-binding site (HBS) deletion mutant (NASP–ΔHBS). The purpose of the current study was to identify possible NASP-associated proteins in HeLa cell nuclei that could elucidate NASP's influence on the cell cycle and chromatin remodeling. For this purpose, we employed a new approach: mass spectrometry identification of initially cross-linked proteins after their separation in a second dimension by reducing SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Of the twelve proteins identified, three appear to be relevant to NASP's function: heat shock protein 90 (HSP90), DNA-activated protein kinase, and ATP-dependent DNA helicase II (70-kDa subunit). Individual protein–protein interactions were tested by immunoprecipitation techniques. This new method can be used for expedited identification of binding partners of different proteins in enriched fractions and as a complementary or alternative strategy to the yeast two-hybrid system and immunoprecipitation methods
Details
- Title: Subtitle
- Mass spectrometry identification of NASP binding partners in HeLa cells
- Creators
- Oleg M. Alekseev - University of North Carolina at Chapel HillRichard T. Richardson - University of North Carolina at Chapel HillMarshall R. Pope - University of North Carolina at Chapel HillMichael G. O'Rand - University of North Carolina at Chapel Hill
- Resource Type
- Journal article
- Publication Details
- Proteins, structure, function, and bioinformatics, Vol.61(1), pp.1-5
- Publisher
- Wiley Subscription Services, Inc., A Wiley Company
- DOI
- 10.1002/prot.20600
- PMID
- 16080155
- ISSN
- 0887-3585
- eISSN
- 1097-0134
- Number of pages
- 5
- Language
- English
- Date published
- 10/01/2005
- Academic Unit
- Medicine Administration
- Record Identifier
- 9984627184002771
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