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

• Published in: Protein expression and purification Vol. 71; no. 1; pp. 103 - 107
• Main Authors: Han, Byeong-Gu, Cho, Jea-Won, Cho, Young-Doo, Kim, Soo-Youl, Yoon, Hye-Jin, Song, Hyun Kyu, Cheong, Hae-Kap, Jeon, Young-Ho, Lee, Dong-ki, Lee, Sangho, Lee, Byung Il
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.05.2010
• Subjects: Amino Acid Sequence; Biochemistry - methods; HRASLS3; HREV107; 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; RARRES3; Receptors, Retinoic Acid - chemistry; Receptors, Retinoic Acid - isolation & purification; Receptors, Retinoic Acid - metabolism; Recombinant Proteins - chemistry; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; RIG1; TIG3; Tumor Suppressor Proteins - chemistry; Tumor Suppressor Proteins - isolation & purification; Tumor Suppressor Proteins - metabolism; HRASLS3; RARRES3; TIG3; RIG1; HREV107
• ISSN: 1046-5928; 1096-0279
• DOI: 10.1016/j.pep.2010.01.018

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 A2 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.