Short Report
On the cytotoxicity of HCR-NTPase in the neuroblastoma cell line SH-SY5Y
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* Corresponding author: Michael Kaufmann mika@uni-wh.de
1 The Protein Chemistry Group, Witten/Herdecke University, Stockumer Str 10, 58453 Witten, Germany
2 Institute for Neurobiochemistry, Witten/Herdecke University, Stockumer Str 10, 58453 Witten, Germany
3 Dept. of Cell Biology, Bielefeld University, Universitätsstr 25, 33501 Bielefeld, Germany
BMC Research Notes 2009, 2:102 doi:10.1186/1756-0500-2-102
Published: 11 June 2009Abstract
Background
The human cancer-related nucleoside triphosphatase (HCR-NTPase) is overexpressed in several tumour tissues including neuroblastoma. HCR-NTPase is an enzyme exhibiting a slow in vitro activity in hydrolysing nucleosidetriphosphates. However, its in vivo function is still unknown. To learn more about the physiological role of HCR-NTPase, we both overexpressed and silenced it in the neuroblastoma cell line SH-SY5Y.
Findings
No effect was observed when the expression of endogenously expressed HCR-NTPase in the cells was silenced by RNA interference. On the other hand, overexpression of HCR-NTPase led to cytotoxicity of the protein in SH-SY5Y cells. Even if the catalytic essential amino acid glutamate 114 was replaced by alanine (E114A-HCR-NTPase), the protein remained cytotoxic. The results could be confirmed by successfully rescuing the cells via RNA interference.
Conclusion
Although expressed in several tumours, at least in SH-SY5Y, HCR-NTPase is not essential for the cells to survive. Increased levels of the protein lead to cytotoxicity due to physical intracellular interactions rather than hydrolysis of nucleosidetriphosphates by its intrinsic residual enzymatic activity.