OBTENCIÓN DE UN SUB-PROTEOMA DE CITOPLASMA DE UNA LÍNEA CELULAR DE TROFOBLASTO MEDIANTE FRACCIONAMIENTO CON DETERGENTES

Sandra Susana Novoa-Herrán; Myriam Sánchez-Gómez

doi:10.18257/raccefyn.35(136).2011.2507

Vol. 35 No. 136 (2011), Chemical Sciences

Vol. 35 No. 136 (2011)

OBTENTION OF CYTOPLASM SUB-PROTEOME OF A TROPHOBLAST CELL LINE BY DETERGENT FRACTIONATION

Chemical Sciences

https://doi.org/10.18257/raccefyn.35(136).2011.2507

Published 2023-12-22

Sandra Susana Novoa-Herrán⁺⁻
Myriam Sánchez-Gómez⁺⁻

Sandra Susana Novoa-Herrán

Estudiante de doctorado.

Myriam Sánchez-Gómez

Profesor Titular. Grupo de Investigación en Hormonas, Departamento de Química, Universidad Nacional de Colombia. Cra. 30 45-03, Bogotá D.C., Colombia.

PDF (Español (España))

How to Cite

Novoa-Herrán, S. S., & Sánchez-Gómez, M. (2023). OBTENTION OF CYTOPLASM SUB-PROTEOME OF A TROPHOBLAST CELL LINE BY DETERGENT FRACTIONATION. Revista De La Academia Colombiana De Ciencias Exactas, Físicas Y Naturales, 35(136), 277–286. https://doi.org/10.18257/raccefyn.35(136).2011.2507

Downloads

Download data is not yet available.

Métricas Alternativas

Dimensions

Abstract

The differential detergent fractionation method described by Rambsy and Makowski in 1999, was modified using a digitonin (0.015%) buffer to permeate HTR8/SVneo human trophoblast cells and obtain the cytoplasm proteins in the soluble fraction, while membrane proteins were dissolved from the pellet with Triton X-100 (0.5%) buffer. We obtained a reproducible membrane-free cytoplasm protein extract suitable for 2-DE, by chloroform/methanol precipitation. In conclusion, the method here described is appropriate for the separation of cytoplasm sub-proteomes from cell cultures, with an optimal reproducibility and recovery to perform proteomic profile analysis by bidimensional electrophoresis.

https://doi.org/10.18257/raccefyn.35(136).2011.2507

Keywords

PDF (Español (España))

References

Dowling, P. M., P. Dowd, A. Henry, M. Glynn, S. Clynes, M. 2007. “Proteomic analysis of isolated membrane fractions from superinvasive cancer cells.” Biochimica et Biophysica Acta 1774:93-101.

Everberg, H., T. Leiding, et al. 2006. “Efficient and non-denaturing membrane solubilization combined with enrichment of membrane protein complexes by detergent/polymer aqueous two-phase partitioning for proteome analysis.” Journal of Chromatography A 1122(1-2): 35-46.

Ferretti, C., L. Bruni, et al. (2007). “Molecular circuits shared by placental and cancer cells, and their implications in the proliferative, invasive and migratory capacities of trophoblasts.” Hum Reprod Update 13(2):121-141.

Foster, L. J., et al. 2005. “Differential expression profiling of membrane proteins by quantitative proteomics in a human mesenchymal stem cell line undergoing osteoblast differentiation”. Stem Cells 23(9):1367-1377.

Gonnet, F., et al. 2003. “MALDI/MS peptide mass fingerprinting for proteome analysis: identification of hydrophobic proteins attached to eucaryote keratinocyte cytoplasmic membrane using different matrices in concert”. Proteome Science 1(1):2.

Graham, C. H., T. S. Hawley, et al. 1993. “Establishment and Characterization of First Trimester Human Trophoblast Cells with Extended Lifespan”. Experimental Cell Research 206(2):204-211.

Jethwaney, D., et al. (2007). “Proteomic analysis of plasma membrane and secretory vesicles from human neutrophils”. Proteome Science 5(1):12.

Liang, X., et al. 2006. “Quantification of membrane and membrane-bound proteins in normal and malignant breast cancer cells isolated from the same patient with primary breast carcinoma.” J. Proteome Res 5(10):2632-2641.

McCarthy, F. M., S. C. Burgess, et al. 2005. “Differential Detergent Fractionation for Non-electrophoretic Eukaryote Cell Proteomics.” Journal of Proteome Research 4(2):316-324.

Payrastre, B., M. Plantavid, et al. 1988. “Characterization of plasma membranes from A431 cells, isolated by self-generating Percoll gradient: a rapid isolation procedure to obtain plasma membranes with functional epidermal growth factor receptors”. Biochimica et Biophysica Acta (BBA) - Biomembranes 939(2):355-365.

Reiner Westermeier, T. N., Hans-Rudolf Höpker 2008. Proteomics in Practice: A Guide to Successful Experimental Design. Weinheim, Wiley-VCH.

Roobol-Bóza, M., V. Dolby, et al. 2004. “Membrane protein isolation by in situ solubilization, partitioning and affinity adsorption in aqueous two-phase systems: Purification of the human type 1 11[beta]-hydroxysteroid dehydrogenase.” Journal of Chromatography A 1043(2):217-223.

Selinsky, B. S. (Ed.) 2003. Membrane Protein Protocols: Expression, Purification, and Characterization. Methods in Molecular Biology, Humana Press.

Vertommen, A., B. Panis, et al. 2011. “Challenges and solutions for the identification of membrane proteins in non-model plants.” Journal of Proteomics 74(8):1165-1181.

Walker, J. M., (Ed.) 2005. The Proteomics Protocols Handbook. Totowa, New Jersey, Humana Press Inc.

Zhang, L., et al. 2008. “Dataset of the plasma membrane proteome of nasopharyngeal carcinoma cell line HNE1 for uncovering protein function.” Acta Biochimica et Biophysica Sinica 40(1):55-70.

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.