Expression, purification and biochemical characterization of the N-terminal regions of human TIG3 and HRASLS3 proteins

Protein Expr Purif. 2010 May;71(1):103-7. doi: 10.1016/j.pep.2010.01.018. Epub 2010 Jan 25.

Abstract

Tarzarotene-induced gene 3 (TIG3) and HRAS-like suppressor (HRASLS3) are members of the HREV107 family of class II tumor suppressors, which are down-regulated in various cancer cells. TIG3 and HRASLS3 also exhibit phospholipase activities. Both proteins share a common domain architecture with hydrophilic N-terminal and hydrophobic C-terminal regions. The hydrophobic C-terminal region is important for tumor suppression. However, the function of the hydrophilic N-terminal region remains elusive. To facilitate biochemical characterizations of TIG3 and HRASLS3, we expressed and purified the N-terminal regions of TIG3 and HRASLS3, designated TIG3 (1-134) and HRASLS3 (1-133), in a bacterial system. We found that the N-terminal regions of TIG3 and HRASLS3 have calcium-independent phospholipase A(2) activities. Limited proteolysis revealed that TIG3 (1-132) is a structural domain in the N-terminal region of TIG3. Our data suggest that the hydrophobic C-terminal regions might be crucial for cellular localization, while the hydrophilic N-terminal regions are sufficient for the enzymatic activity of both TIG3 and HRASLS3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Biochemistry / methods*
  • Humans
  • Intracellular Signaling Peptides and Proteins / chemistry*
  • Intracellular Signaling Peptides and Proteins / isolation & purification*
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Molecular Sequence Data
  • Phospholipases A2 / metabolism
  • Phospholipases A2, Calcium-Independent
  • Protein Structure, Tertiary
  • Receptors, Retinoic Acid / chemistry*
  • Receptors, Retinoic Acid / isolation & purification*
  • Receptors, Retinoic Acid / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Tumor Suppressor Proteins / chemistry*
  • Tumor Suppressor Proteins / isolation & purification*
  • Tumor Suppressor Proteins / metabolism

Substances

  • Intracellular Signaling Peptides and Proteins
  • PLAAT4 protein, human
  • Receptors, Retinoic Acid
  • Recombinant Proteins
  • Tumor Suppressor Proteins
  • PLAAT3 protein, human
  • Phospholipases A2
  • Phospholipases A2, Calcium-Independent