Reactivity of glutaredoxins 1, 2 and 3 from Escherichia coli and protein disulfide isomerase towards glutathionyl‐mixed disulfides in ribonuclease A

FEBS Letters

Åslund, Fredrik²  Vlamis-Gardikas, Alexios¹  Lundström-Ljung, Johanna¹  Holmgren, Arne¹ 

[1] Medical Nobel Institute of Biochemistry, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 77 Stockholm, Sweden;Department of Microbiology and Molecular Genetics, Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115, USA

关键词: Glutaredoxin;    Protein disulfide isomerase;    Glutathione;    Endoplasmic reticulum;    Mixed disulfide;    DTT;    dithiothreitol;    ES-MS;    electrospray mass spectrometry;    Grx 1;    2 and 3;    glutaredoxin 1;    2 and 3 from Escherichia coli;    GS residue;    glutathionyl residue;    GuHCl;    guanidinium hydrochloride;    PDI;    bovine protein disulfide isomerase;    RNase;    bovine pancreatic ribonuclease A type III;    RNase-SG;    ribonuclease A derivatized with glutathione;    RNase-SH;    fully reduced ribonuclease A;   

DOI  :  10.1016/S0014-5793(98)01698-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have examined the activity of protein disulfide isomerase (PDI) and glutaredoxin (Grx) 1, 2 and 3 from Escherichia coli to catalyze the cleavage of glutathionylated ribonuclease A (RNase-SG) by 1 mM GSH to yield reduced RNase. Apparent K _m values for RNase-SG were similar, 2–10 μM, for Grx 1, 3 and PDI but Grx 1 and Grx 3 showed 500-fold higher turnover numbers than PDI. The atypical Grx 2 also catalyzed deglutathionylation by GSH, but had higher K _m and apparent turnover number values compared to the two classical Grx. Refolding of RNase in a glutathione redox buffer was catalyzed by PDI. However, it could be measured only after a characteristic lag phase that was shortened by all three E. coli Grxs in a concentration-dependent manner. A role of the glutaredoxin mechanism in the endoplasmic reticulum is suggested.

【 授权许可】

Unknown