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Issue 5, 2009
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Proteomic profiling of the stem cellresponse to retinoic acid and synthetic retinoid analogues: identification of major retinoid-inducible proteins

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Abstract

The natural retinoid, all-trans retinoic acid (ATRA), is widely used to direct the in vitro differentiation of stem cells. However, substantial degradation and isomerisation of ATRA in response to UV-vis light has serious implications with regard to experimental reproducibility and standardisation. We present the novel application of proteomic biomarker profiling technology to stem cell lysates to rapidly compare the differentiation effects of ATRA with those of two stable synthetic retinoid analogues, EC19 and EC23, which have both been shown to induce differentiation in the embryonal carcinoma cell line TERA2.cl.SP12. MALDI-TOF MS (matrix-assisted laser desorption ionisation time-of-flight mass spectrometry) protein profiles support previous findings into the functional relationships between these compounds in the TERA2.cl.SP12 line. Subsequent analysis of protein peak data enabled the semi-quantitative comparison of individual retinoid-responsive proteins. We have used ion exchange chromatographicprotein separation to enrich for retinoid-inducible proteins, thereby facilitating their identification from SDS-PAGE gels. The cellular retinoid-responsive proteins CRABP-I, CRABP-II, and CRBP-I were up-regulated in response to ATRA and EC23, indicating a bona fideretinoid pathway response to the synthetic compound. In addition, the actin filament regulatory protein profilin-1 and the microtubule regulator stathmin were also elevated following treatment with both ATRA and EC23. The up-regulation of profilin-1 and stathmin associated with retinoid-induced neural differentiation correlates with their known roles in cytoskeletal reorganisation during axonal development. Immunological analysis via western blotting confirmed the identification of CRABP-I, profilin-1 and stathmin, and supported their observed regulation in response to the retinoid treatments.

Graphical abstract: Proteomic profiling of the stem cellresponse to retinoic acid and synthetic retinoid analogues: identification of major retinoid-inducible proteins

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Publication details

The article was received on 14 Oct 2008, accepted on 11 Feb 2009 and first published on 12 Mar 2009


Article type: Paper
DOI: 10.1039/B817912C
Mol. BioSyst., 2009,5, 458-471

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    Proteomic profiling of the stem cellresponse to retinoic acid and synthetic retinoid analogues: identification of major retinoid-inducible proteins

    D. J. Maltman, V. B. Christie, J. C. Collings, J. H. Barnard, S. Fenyk, T. B. Marder, A. Whiting and S. A. Przyborski, Mol. BioSyst., 2009, 5, 458
    DOI: 10.1039/B817912C

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