Pharmaceutical Sciences Faculty Publications

Probing Early Growth Response 1 Interacting Proteins at the Active Promoter in Osteoblast Cells Using Oligoprecipitation and Mass Spectrometry

Document Type

Article

Publication Date

8-1-2006

Journal Title

Journal of Proteome Research

ISSN

1535-3893

Volume

5

Issue

8

First Page

1931

Last Page

1939

DOI

10.1021/pr060009l

PubMed ID

16889415

Abstract

Current advances in proteomics have allowed for a rapidly expanding integration of associated methodologies with more traditional molecular and biochemical approaches to the study of cell function. Recent studies on the role of inorganic phosphate have suggested this ion is a novel signaling molecule capable of altering the function of numerous cell types. Elevated inorganic phosphate generated in the extracellular microenvironment by differentiating osteoblasts has recently been determined to act through a largely uncharacterized mechanism as an important signaling molecule responsible for altering the transcription of various genes during osteoblast differentiation. The transcription factor, early growth response protein 1 (EGR1), has previously been shown to be involved in the early response of osteoblasts to inorganic phosphate. To elucidate the role of EGR1 as a potential early regulator of transcription in the inorganic phosphate response, an oligoprecipitation procedure was optimized to capture the DNA bound, transcriptionally active form of EGR1. The interacting proteins thusly captured were identified using mass spectrometry (MS). Proteins involved in transcription, RNA processing, and chromatin modification were identified by this approach. The combined oligoprecipitation-MS approach presented here is highly effective for isolating and characterizing entire transcriptional complexes in the DNA bound state and is broadly extendable to the identification of both known and unknown transcription factor protein complexes.

Keywords

Early growth response protein 1, mass spectrometry, molecular sequence data, osteoblasts, phosphates, promoter regions, genetic, proteome, transcription, genetic

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