[Affinity Detection and Quantification of Receptors of TRAIL Based on Their Thermostability].

Abstract

OBJECTIVE:

To develop a way for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptors quantification in cancer via its' thermostability.

METHODS:

Endougenous alkaline phosphatase (AP) activity and denaturation temperature of pancreatic cancer cell lines AsPC-1 and Capan-2 were detected. Boiling treated recombinant protein death receptor 5 (DR5), named DR5 AP, as well as pancreatic cancer cells lines AsPC-1 and Capan-2 were incubated with AP-tagged TRAIL (AP-TRAIL), and then reacted with Reagent A and Reagent S, the substrate of AP, to quantitive and in site detection of the receptor.

RESULTS:

The endougenous AP activity of pancreatic cancer cells lines AsPC-1 and Capan-2 could not be totally inactivated by incubated at 65 °C, thus inhibited the detection of TRAIL receptor, but the activity was dramatically decreased after treated with boiling water, whereas the DR5-AP was thermal stable. The surface receptor of AsPC-1 and Capan-2 could be recognized and bound by AP-TRAIL after treated at 100 °C, the readings were 2. 210±0. 393 and 2. 027±0. 019.

CONCLUSION:

The TRAIL receptors are thermostable and this may provide a better diagnosis and prognosis of cancer as well as personalize cancer therapy.

• Authors: Yang Xu, Hai-peng Liu, Tian-zhen Long, Guang-ping Luo, Peng Liang
• Journal: Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition (Sichuan Da Xue Xue Bao Yi Xue Ban) Vol. 46 Issue 3 Pg. 449-52 (May 2015) ISSN: 1672-173X [Print] China
• PMID: 26121872 (Publication Type: Journal Article)

Chemical References

• Receptors, TNF-Related Apoptosis-Inducing Ligand
• Recombinant Proteins

Topics

• Cell Line, Tumor
• Hot Temperature
• Humans
• Pancreatic Neoplasms (metabolism)
• Protein Stability
• Receptors, TNF-Related Apoptosis-Inducing Ligand (metabolism)
• Recombinant Proteins (metabolism)