Targeted label-free quantification of interleukin-8 in PMA-activated U937 cell secretome by nanoLC-ESI-MS/MS-sSRM.
Emirbayer PE, Gerer K, Hoyer S, Pischetsrieder M (2017)
Publication Status: Published
Publication Type: Journal article, Original article
Publication year: 2017
Journal
Abstract
Monocytes are a part of the innate immune system. Their differentiation into macrophages changes their cellular proteome and secretome. Particularly secretome components such as cytokines are crucial for immune response and inflammation in many diseases. Differentiation of human lymphoma cell line U937 can be triggered by phorbol 12-myristate 13-acetate (PMA). Screening of the cytokine release in U937 upon PMA stimulation by cytometric bead array almost exclusively showed interleukin-8. Next, a label-free nanoLC-ESI-MS/MS-sSRM method for quantification of interleukin-8 in the cell secretome was established and applied to monitor the time kinetics of PMA treatment in different concentrations. Targeted secretome analysis was achieved by scheduled SRM-MS using one proteotypic peptide as precursor ion and four mass transitions. Label-free quantification was performed by external calibration using interleukin-8 standard. Validation results indicated that the method was suited for the quantification of interleukin-8 in the secretome. The maximal interleukin-8 release of 62.4 ng/mL was observed after incubating cells treated by 50 ng/mL PMA for 48 h. The method can now be used for quantification of interleukin-8 release from different cells under various conditions. Furthermore, it can be easily expanded to other secreted proteins detected by untargeted screening methods. This article is protected by copyright. All rights reserved.
Authors with CRIS profile
Pelin Esma Emirbayer
Lehrstuhl für Lebensmittelchemie (Henriette-Schmidt-Burkhardt Lehrstuhl)
Stefanie Hoyer
Department of Dermatology
Monika Pischetsrieder
Lehrstuhl für Lebensmittelchemie (Henriette-Schmidt-Burkhardt Lehrstuhl)
Additional Organisation(s)
Involved external institutions
Germany (DE)How to cite
APA:
Emirbayer, P.E., Gerer, K., Hoyer, S., & Pischetsrieder, M. (2017). Targeted label-free quantification of interleukin-8 in PMA-activated U937 cell secretome by nanoLC-ESI-MS/MS-sSRM. Proteomics. https://dx.doi.org/10.1002/pmic.201600455
MLA:
Emirbayer, Pelin Esma, et al. "Targeted label-free quantification of interleukin-8 in PMA-activated U937 cell secretome by nanoLC-ESI-MS/MS-sSRM." Proteomics (2017).
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