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A proteomic approach for dissecting H-Ras signaling networks in NIH/3T3 mouse embryonic fibroblast cells.

Publication ,  Journal Article
Park, JW; Kim, S; Bahk, YY
Published in: Proteomics
April 2006

To elucidate an understanding into H-Ras protein network, we have established various oncogene H-Ras-expressing NIH/3T3 mouse embryonic fibroblast cell clones, which are expressing G12V H-Ras, G12R H-Ras, and G12V/T35S H-Ras proteins under the tight control of expression by an antibiotic doxycycline. Here we provide a catalog of proteome profiles in total cell lysate derived from the oncogenic and partial loss of function H-Ras-expressing NIH/3T3 cells. In this biological context, we compared total proteome changes by the combined methods of 2-DE, quantitative image analysis and MALDI-TOF-MS analysis both commonly in oncogenic and partial loss of function H-Ras expression system. Thus, we tried to dissect H-Ras signaling pathway, especially a downstream effector molecule, Raf in NIH/3T3 cells using proteomics tools. In this study, we centralized upon the proteome profile changes as common targets for oncogenic H-Ras and a partial loss of function H-Ras in the H-Ras-expressing cells. Thirteen protein spots were selected as what the staining intensities on the gels for 2-DE images from both kinds of cells were consistently changed in their protein expression level. Differentially regulated expression was further confirmed for some subsets of candidates by semiquantitative RT-PCR and Western blot analysis using specific antibodies. Taken together, our results obtained and present here show that the comparative analysis of proteome from oncogenic and partial loss of function H-Ras-expressing cells has yielded interpretable data to elucidate the protein network directly and/or indirectly.

Duke Scholars

Published In

Proteomics

DOI

ISSN

1615-9853

Publication Date

April 2006

Volume

6

Issue

8

Start / End Page

2433 / 2443

Location

Germany

Related Subject Headings

  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Signal Transduction
  • Proto-Oncogene Proteins p21(ras)
  • Proteomics
  • NIH 3T3 Cells
  • Mice
  • Fibroblasts
  • Electrophoresis, Gel, Two-Dimensional
  • Doxycycline
  • Biochemistry & Molecular Biology
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Park, J. W., Kim, S., & Bahk, Y. Y. (2006). A proteomic approach for dissecting H-Ras signaling networks in NIH/3T3 mouse embryonic fibroblast cells. Proteomics, 6(8), 2433–2443. https://doi.org/10.1002/pmic.200500688
Park, Jung Wook, Seyoon Kim, and Young Yil Bahk. “A proteomic approach for dissecting H-Ras signaling networks in NIH/3T3 mouse embryonic fibroblast cells.Proteomics 6, no. 8 (April 2006): 2433–43. https://doi.org/10.1002/pmic.200500688.
Park, Jung Wook, et al. “A proteomic approach for dissecting H-Ras signaling networks in NIH/3T3 mouse embryonic fibroblast cells.Proteomics, vol. 6, no. 8, Apr. 2006, pp. 2433–43. Pubmed, doi:10.1002/pmic.200500688.
Journal cover image

Published In

Proteomics

DOI

ISSN

1615-9853

Publication Date

April 2006

Volume

6

Issue

8

Start / End Page

2433 / 2443

Location

Germany

Related Subject Headings

  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Signal Transduction
  • Proto-Oncogene Proteins p21(ras)
  • Proteomics
  • NIH 3T3 Cells
  • Mice
  • Fibroblasts
  • Electrophoresis, Gel, Two-Dimensional
  • Doxycycline
  • Biochemistry & Molecular Biology