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Related Experiment Videos

Protein expression in a transformed trabecular meshwork cell line: proteome analysis.

H Thomas Steely1, Glen W Dillow, Liangqian Bian

  • 1Alcon Research, Ltd., Fort Worth, TX 76134, USA. thomas.steely@alconlabs.com

Molecular Vision
|April 26, 2006
PubMed
Summary

This study initiated a proteomic survey of human trabecular meshwork (TM) cells, identifying 87 primary proteins and 93 variants. This research provides a foundation for understanding TM proteome in normal and glaucomatous conditions.

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Area of Science:

  • Proteomics
  • Human physiology
  • Ocular biology

Background:

  • Characterizing the human trabecular meshwork (TM) proteome is challenging due to limited tissue availability and slow cell culture growth.
  • A transformed TM cell line (GTM3) was previously established, exhibiting similar protein expression and signaling to normal cells.

Purpose of the Study:

  • To initiate a proteomic survey of GTM3 cells.
  • To establish a foundation for characterizing the complete human TM proteome.
  • To aid in understanding protein expression in normal and glaucomatous TM.

Main Methods:

  • GTM3 cells were cultured, harvested, and solubilized.
  • Proteins were separated using immobilized isoelectric focusing (IEF) and 10% SDS-PAGE.
  • Protein identification was performed using LC/MS/MS analysis and TurboSequest algorithm against the NCBI human protein database.

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Main Results:

  • Identification of 87 primary proteins and 93 protein variants.
  • Creation of a website (TM proteome) detailing protein data, including gel location, peptide sequences, and physicochemical properties.
  • Comprehensive cataloging of identified proteins and their characteristics.

Conclusions:

  • Proteomic analysis of a transformed human TM cell line was successfully initiated.
  • The generated data will be used comparatively for analyzing normal TM tissue and cultured cells.
  • This study provides a foundational dataset for future research on TM proteome in health and disease.