a depletion by transglutaminase 2 and l-calpain occurs in parallel with the ubiquitin – proteasome pathway

0006-291X/$ see front matter 2010 Elsevier Inc. A doi:10.1016/j.bbrc.2010.07.078 Abbreviations: TGase, transglutaminase; IKK, I-j factor-jB; I-jBa, inhibitory subunit of NF-jB; CTM, c * Corresponding author. ** Corresponding author. E-mail addresses: kimsooyoul@gmail.com (S.-Y. Kim D. Bae). Transglutaminase 2 (TGase2) is a calcium-dependent, cross-linking enzyme that catalyzes iso-peptide bond formation between peptide-bound lysine and glutamine residues. TGase 2 can activate NF-jB through the polymerization-mediated depletion of I-jBa without IKK activation. This NF-jB activation mechanism is associated with drug resistance in cancer cells. However, the polymers cannot be detected in cells, while TGase 2 over-expression depletes free I-jBa, which raises the question of how the polymerized I-jBa can be metabolized in cells. Among proteasome, lysosome and calpain systems, calpain inhibition was found to effectively increase the accumulation of I-jBa polymers in MCF7 cells transfected with TGase 2, and induced high levels of I-jBa polymers as well in MDA-MB-231 breast cancer cells that naturally express a high level of TGase 2. Inhibition of calpain also boosted the level of I-jBa polymers in HEK-293 cells in case of TGase 2 transfection either with I-jBa or I-jBa mutant (S32A, S36A). Interestingly, the combined inhibition of calpain and the proteasome resulted in an increased accumulation of both I-jBa polymers and I-jBa, concurrent with an inhibition of NF-jB activity in MDA-MB-231 cells. This suggests that l-calpain proteasome-dependent I-jBa polymer degradation may contribute to cancer progression through constitutive NF-jB activation. 2010 Elsevier Inc. All rights reserved.

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