Phosphopeptide mapping of proteins ectopically expressed in tissue culture cell lines

Beth A. Firulli, David M. Virshup, Anthony B. Firulli

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Post-translational modifications such as phosphorylation play a vital role in the regulation of protein function. In our study of the basic Helix-loop-Helix (bHLH) transcription factor HAND1, it was suspected that HAND1 was being phosphorylated during trophoblast giant cell differentiation and that coexpression of a constitutively active kinase with HAND1 resulted in changes in the proteins dimerization profile. In order to accurately document HAND1 phosphorylation and identify the resides being modified, we employed metabolic cell labeling with 32P of tissue culture cells coexpressing a Flag-epitope tagged HAND1 along with a number of active kinases and phosphatase subunits. We generated phosphopeptide maps of the phosphorylated HAND1 using the methods described below and linked these modifications to changes in HAND1 biological function.

Original languageEnglish (US)
Pages (from-to)16-22
Number of pages7
JournalBiological Procedures Online
Volume6
Issue number1
DOIs
StatePublished - Jan 15 2004

Fingerprint

Phosphopeptides
Tissue culture
Phosphorylation
Phosphotransferases
Protein Multimerization
Basic Helix-Loop-Helix Transcription Factors
Cell Line
Dimerization
Trophoblasts
Giant Cells
Post Translational Protein Processing
Phosphoric Monoester Hydrolases
Labeling
Epitopes
Cell Differentiation
Proteins
Cell Culture Techniques

Keywords

  • Dimerization
  • Helix-loop-Helix mofits
  • Phosphorylation

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Phosphopeptide mapping of proteins ectopically expressed in tissue culture cell lines. / Firulli, Beth A.; Virshup, David M.; Firulli, Anthony B.

In: Biological Procedures Online, Vol. 6, No. 1, 15.01.2004, p. 16-22.

Research output: Contribution to journalArticle

@article{d9cbe58b598644a6b454faa8fad288cd,
title = "Phosphopeptide mapping of proteins ectopically expressed in tissue culture cell lines",
abstract = "Post-translational modifications such as phosphorylation play a vital role in the regulation of protein function. In our study of the basic Helix-loop-Helix (bHLH) transcription factor HAND1, it was suspected that HAND1 was being phosphorylated during trophoblast giant cell differentiation and that coexpression of a constitutively active kinase with HAND1 resulted in changes in the proteins dimerization profile. In order to accurately document HAND1 phosphorylation and identify the resides being modified, we employed metabolic cell labeling with 32P of tissue culture cells coexpressing a Flag-epitope tagged HAND1 along with a number of active kinases and phosphatase subunits. We generated phosphopeptide maps of the phosphorylated HAND1 using the methods described below and linked these modifications to changes in HAND1 biological function.",
keywords = "Dimerization, Helix-loop-Helix mofits, Phosphorylation",
author = "Firulli, {Beth A.} and Virshup, {David M.} and Firulli, {Anthony B.}",
year = "2004",
month = "1",
day = "15",
doi = "10.1251/bpo69",
language = "English (US)",
volume = "6",
pages = "16--22",
journal = "Biological Procedures Online",
issn = "1480-9222",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - Phosphopeptide mapping of proteins ectopically expressed in tissue culture cell lines

AU - Firulli, Beth A.

AU - Virshup, David M.

AU - Firulli, Anthony B.

PY - 2004/1/15

Y1 - 2004/1/15

N2 - Post-translational modifications such as phosphorylation play a vital role in the regulation of protein function. In our study of the basic Helix-loop-Helix (bHLH) transcription factor HAND1, it was suspected that HAND1 was being phosphorylated during trophoblast giant cell differentiation and that coexpression of a constitutively active kinase with HAND1 resulted in changes in the proteins dimerization profile. In order to accurately document HAND1 phosphorylation and identify the resides being modified, we employed metabolic cell labeling with 32P of tissue culture cells coexpressing a Flag-epitope tagged HAND1 along with a number of active kinases and phosphatase subunits. We generated phosphopeptide maps of the phosphorylated HAND1 using the methods described below and linked these modifications to changes in HAND1 biological function.

AB - Post-translational modifications such as phosphorylation play a vital role in the regulation of protein function. In our study of the basic Helix-loop-Helix (bHLH) transcription factor HAND1, it was suspected that HAND1 was being phosphorylated during trophoblast giant cell differentiation and that coexpression of a constitutively active kinase with HAND1 resulted in changes in the proteins dimerization profile. In order to accurately document HAND1 phosphorylation and identify the resides being modified, we employed metabolic cell labeling with 32P of tissue culture cells coexpressing a Flag-epitope tagged HAND1 along with a number of active kinases and phosphatase subunits. We generated phosphopeptide maps of the phosphorylated HAND1 using the methods described below and linked these modifications to changes in HAND1 biological function.

KW - Dimerization

KW - Helix-loop-Helix mofits

KW - Phosphorylation

UR - http://www.scopus.com/inward/record.url?scp=2342499883&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2342499883&partnerID=8YFLogxK

U2 - 10.1251/bpo69

DO - 10.1251/bpo69

M3 - Article

AN - SCOPUS:2342499883

VL - 6

SP - 16

EP - 22

JO - Biological Procedures Online

JF - Biological Procedures Online

SN - 1480-9222

IS - 1

ER -