The threonine protease activity of testes-specific protease 50 (TSP50) is essential for its function in cell proliferation.

PloS One
Yu-Yin LiYu-Xin Li

Abstract

Testes-specific protease 50 (TSP50), a newly discovered threonine enzyme, has similar amino acid sequences and enzymatic structures to those of many serine proteases. It may be an oncogene. TSP50 is up-regulated in breast cancer epithelial cells, and ectopic expression of TSP50 in TSP50-deficient Chinese hamster ovary (CHO) cells has been found to promote cell proliferation. However, the mechanisms by which TSP50 exerts its growth-promoting effects are not yet fully understood. To delineate whether the threonine protease activity of TSP50 is essential to its function in cell proliferation, we constructed and characterized a mutant TSP50, called TSP50 T310A, which was identified as a protease-dead mutant of TSP50. By a series of proliferation analyses, colony formation assays and apoptosis analyses, we showed that T310A mutation significantly depresses TSP50-induced cell proliferation in vitro. Next, the CHO stable cell line expressing either wild-type or T310A mutant TSP50 was injected subcutaneously into nude mice. We found that the T310A mutation could abolish the tumorigenicity of TSP50 in vivo. A mechanism investigation revealed that the T310A mutation prevented interaction between TSP50 and the NF-κBIκBα complex, which is ...Continue Reading

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Citations

May 7, 2014·Gastroenterology Research and Practice·Fang LiuRongcheng Luo
Aug 1, 2014·Tumour Biology : the Journal of the International Society for Oncodevelopmental Biology and Medicine·Xiaopeng Zhang
Dec 18, 2015·Cell Biology International·Yu-Le YongHuijie Bian
Oct 27, 2016·Tumour Biology : the Journal of the International Society for Oncodevelopmental Biology and Medicine·Danfeng WangYuxin Li

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Methods Mentioned

BETA
confocal microscopy
PCR
Flow Cytometry
Assay
PMA
immunoprecipitation
co-immunoprecipitation assay
nuclear translocation
transfection
electrophoresis

Software Mentioned

pred
THREADER
PyMOL
HH
blast
MODEL
ImageJ
TASSER

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