Email this article Human SLURP-1 and SLURP-2 Proteins Acting on Nicotinic Acetylcholine Receptors Reduce Proliferation of Human Colorectal Adenocarcinoma HT-29 Cells
- Authors: Lyukmanova E.N.1,2, Shulepko M.A.1,2, Bychkov M.L.1,2, Shenkarev Z.O.1,2, Paramonov A.S.1,2, Chugunov A.O.1,2, Arseniev A.S.1,3, Dolgikh D.A.1,2, Kirpichnikov M.P.1,2
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Affiliations:
- Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry
- Lomonosov Moscow State University
- Moscow Institute of Physics and Technology (State University)
- Issue: Vol 6, No 4 (2014)
- Pages: 60-66
- Section: Research Articles
- URL: https://actanaturae.ru/2075-8251/article/view/10526
- DOI: https://doi.org/10.32607/20758251-2014-6-4-60-66
- ID: 10526
Cite item
Abstract
Human secreted Ly-6/uPAR related proteins (SLURP-1 and SLURP-2) are produced by various cells, including the epithelium and immune system. These proteins act as autocrine/paracrine hormones regulating the growth and differentiation of keratinocytes and are also involved in the control of inflammation and malignant cell transformation. These effects are assumed to be mediated by the interactions of SLURP-1 and SLURP-2 with the α7 and α3β2 subtypes of nicotinic acetylcholine receptors (nAChRs), respectively. Available knowledge about the molecular mechanism underling the SLURP-1 and SLURP-2 effects is very limited. SLURP-2 remains one of the most poorly studied proteins of the Ly-6/uPAR family. In this study, we designed for the first time a bacterial system for SLURP-2 expression and a protocol for refolding of the protein from cytoplasmic inclusion bodies. Milligram quantities of recombinant SLURP-2 and its 13С-15N-labeled analog were obtained. The recombinant protein was characterized by NMR spectroscopy, and a structural model was developed. A comparative study of the SLURP-1 and SLURP-2 effects on the epithelial cell growth was conducted using human colorectal adenocarcinoma HT-29 cells, which express only α7-nAChRs. A pronounced antiproliferative effect of both proteins was observed. Incubation of cells with 1 μM SLURP-1 and 1 μM SLURP-2 during 48 h led to a reduction in the cell number down to ~ 54 and 63% relative to the control, respectively. Fluorescent microscopy did not reveal either apoptotic or necrotic cell death. An analysis of the dose-response curve revealed the concentration-dependent mode of the SLURP-1 and SLURP-2 action with EC50 ~ 0.1 and 0.2 nM, respectively. These findings suggest that the α7-nAChR is the main receptor responsible for the antiproliferative effect of SLURP proteins in epithelial cells.
About the authors
E. N. Lyukmanova
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Author for correspondence.
Email: ekaterina-lyukmanova@yandex.ru
Russian Federation
M. A. Shulepko
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: ekaterina-lyukmanova@yandex.ru
Russian Federation
M. L. Bychkov
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: ekaterina-lyukmanova@yandex.ru
Russian Federation
Z. O. Shenkarev
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: ekaterina-lyukmanova@yandex.ru
Russian Federation
A. S. Paramonov
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: ekaterina-lyukmanova@yandex.ru
Russian Federation
A. O. Chugunov
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: ekaterina-lyukmanova@yandex.ru
Russian Federation
A. S. Arseniev
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry; Moscow Institute of Physics and Technology (State University)
Email: ekaterina-lyukmanova@yandex.ru
Russian Federation
D. A. Dolgikh
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: ekaterina-lyukmanova@yandex.ru
Russian Federation
M. P. Kirpichnikov
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: ekaterina-lyukmanova@yandex.ru
Russian Federation
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