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Profiling Optimal Conditions for Capturing EDEM Proteins Complexes in Melanoma Using Mass Spectrometry

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Advancements of Mass Spectrometry in Biomedical Research

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 1140))

Abstract

Endoplasmic reticulum (ER) resident and secretory proteins that fail to reach their native conformation are selected for degradation through the ER-Associated Degradation (ERAD) pathway. The ER degradation-enhancing alpha-mannosidase-like proteins (EDEMs) were shown to be involved in this pathway but their precise role is still under investigation. Mass spectrometry analysis has contributed significantly to the characterization of protein complexes in the last years. The recent advancements in instrumentation, especially within resolution and speed can provide unique insights concerning the molecular architecture of protein-protein interactions in systems biology. Previous reports have suggested that several protein complexes in ERAD are sensitive to the extraction conditions. Indeed, whilst EDEM proteins can be recovered in most detergents, some of their partners are not solubilized, which further emphasizes the importance of the experimental setup. Here, we define such dynamic interactions of EDEM proteins by employing offline protein fractionation, nanoLC-MS/MS and describe how mass spectrometry can contribute to the characterization of such complexes, particularly within a disease context like melanoma.

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Acknowledgments

We acknowledge support for funding to the Romanian Ministry of Research and Innovation, CNCS-UEFISCDI grant PN-III-P1-1.1-PD-2016-1528 and CCCDI-UEFISCDI grant PNCDI-III-PCCDI-2018-1, PN-III-P4-ID-PCE-2016-0650. Part of this work constituted the Ph.D. thesis of Cristian V.A. Munteanu, which received financial support from European Social Fund for Sectorial Operational Program Human Resources Development 2007–2013 grant no. POSDRU/159/1.5/S/135760 and Gabriela N. Chiriţoiu, supported by Romanian Academy. All the authors were partially funded by Romanian Academy.

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Author notes

  1. Cristian V. A. Munteanu and Gabriela N. Chiriţoiu contributed equally to this work.

Authors and Affiliations

  1. Department of Bioinformatics and Structural Biochemistry, Institute of Biochemistry, Bucharest, Romania

    Cristian V. A. Munteanu & Andrei-Jose Petrescu

  2. Department of Molecular Cell Biology, Institute of Biochemistry, Bucharest, Romania

    Gabriela N. Chiriţoiu & Ștefana M. Petrescu

Authors
  1. Cristian V. A. Munteanu
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  2. Gabriela N. Chiriţoiu
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  3. Andrei-Jose Petrescu
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  4. Ștefana M. Petrescu
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Corresponding author

Correspondence to Ștefana M. Petrescu .

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Editors and Affiliations

  1. Biochemistry and Proteomics Group, Department of Chemistry and Biomolecular Science, Clarkson University, Potsdam, NY, USA

    Alisa G. Woods

  2. Biochemistry and Proteomics Group, Department of Chemistry and Biomolecular Science, Clarkson University, Potsdam, NY, USA

    Costel C. Darie

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Munteanu, C.V.A., Chiriţoiu, G.N., Petrescu, AJ., Petrescu, Ș.M. (2019). Profiling Optimal Conditions for Capturing EDEM Proteins Complexes in Melanoma Using Mass Spectrometry. In: Woods, A., Darie, C. (eds) Advancements of Mass Spectrometry in Biomedical Research. Advances in Experimental Medicine and Biology, vol 1140. Springer, Cham. https://doi.org/10.1007/978-3-030-15950-4_9

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