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Purification and properties of LR1, an inducible DNA binding protein from mammalian B lymphocytes

M Williams1, L A Hanakahi, N Maizels

  • 1Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06510.

The Journal of Biological Chemistry
|June 25, 1993
PubMed

Insights

Researchers purified LR1, a lipopolysaccharide-inducible B cell DNA binding protein. Its activity depends on N-acetylglucosamine modification, crucial for binding immunoglobulin gene regions.

Area of Science:

  • Immunology
  • Molecular Biology
  • Biochemistry

Background:

  • LR1 is a B cell-specific DNA binding activity.
  • It binds to sites within the immunoglobulin heavy chain enhancer and switch regions.
  • LR1 is inducible by lipopolysaccharide.

Purpose of the Study:

  • To purify the LR1 protein.
  • To investigate the post-translational modifications of LR1.
  • To understand the regulatory mechanisms of LR1 DNA binding activity.

Main Methods:

  • Purification of LR1 from nuclear extracts of PD31 murine pre-B cells using three chromatographic steps.
  • Analysis of LR1 DNA binding activity.
  • Investigation of post-translational modifications, specifically N-acetylglucosamine addition.
  • Anion exchange chromatography using Mono Q resin.

Main Results:

  • Successfully purified the 106-kDa LR1 protein.
  • Demonstrated that LR1 DNA binding activity is dependent on phosphorylation.
  • Showed that LR1 undergoes post-translational modification by N-acetylglucosamine addition.
  • Observed that DNA binding activity is restricted to a narrow peak at high salt concentrations during anion exchange chromatography, indicating the importance of modification.

Conclusions:

  • LR1 is a lipopolysaccharide-inducible, B cell-specific DNA binding protein purified from murine pre-B cells.
  • Post-translational modification, specifically N-acetylglucosamine addition, is critical for LR1's DNA binding activity.
  • These modifications regulate LR1's function in immunoglobulin gene regulation.

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