Differential incorporation of biotinylated polyamines by transglutaminase 2

FEBS Letters

Kim, Chai-Wan¹  Choi, Kyung-Ho¹  Kwon, Joon-Cheol¹  Jeon, Ju-Hong¹  Kim, Kyu-il²  Kang, Heun-Soo²  Shin, Dong-Myung¹  Cho, Sung-Yup¹  Kim, In-Gyu¹ 

[1] Department of Biochemistry and Molecular Biology/Aging and Apoptosis Research Center (AARC), Seoul National University College of Medicine, 28 Yongon Dong, Chongno Gu, Seoul 110-799, South Korea;Metabolic Engineering Laboratories, Seoul 110-510, South Korea

关键词: Transglutaminase 2;    Biotinylated spermine;    Polyamine conjugation;    Post-translational modification;    TGase;    transglutaminase;    BP;    biotinylated pentylamine;    BS;    biotinylated spermine;    THF;    tetrahydrofuran;    TFA;    trifluoroacetate;    MeOH;    methanol;    EtOAc;    ethylacetate;    DCC;    1;    3-dicyclohexylcarbodiimide;    HOBt;    1-hydroxybenzotriazole;    DMF;    dimethylformamide;    TLC;    thin layer chromatography;   

DOI  :  10.1016/S0014-5793(02)03836-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Polyamine incorporation or cross-linking of proteins, post-translational modifications mediated by transglutaminase 2 (TGase 2), have been implicated in a variety of physiological functions including cell adhesion, extracellular matrix formation and apoptosis. To better understand the intracellular regulation mechanism of TGase 2, the properties of biotinylated polyamines as substrates for determining in situ TGase activity were analyzed. We synthesized biotinylated spermine (BS), and compared the in vitro and in situ incorporation of BS with that of biotinylated pentylamine (BP), which is an artificial polyamine derivative. When measured in vitro, BP showed a significantly higher incorporation rate than BS. In contrast, in situ incorporation of both BS and BP was not detected even in TGase 2-overexpressed 293 cells. Cells exposed to high calcium showed a marked increase of BP incorporation but not of BS. These data indicate that the in situ activity of TGase 2 gives different results with different substrates, and suggest the possibility of overrepresentation of in situ TGase 2 activity when assayed with BP. Therefore, careful interpretation or evaluation of in situ TGase 2 activity may be required.

【 授权许可】

Unknown