AI Article Synopsis
- The study aimed to enhance understanding of chronic inflammatory diseases by identifying newly synthesized proteins induced by the cytokines TNF-α and IL-1β in THP-1 cells.
- Researchers combined two innovative techniques, bio-orthogonal non-canonical amino acid tagging (BONCAT) and iTRAQ-based proteomics, to analyze these nascent proteins.
- This approach successfully revealed the nascent proteome associated with the inflammatory response, providing insights into molecular mechanisms that could lead to new therapeutic strategies.
Article Abstract
An impediment in the development of new therapeutic strategies for chronic inflammatory diseases is the limited understanding of underlying molecular mechanisms. The objective of this study was to identify newly synthesized (nascent) proteins induced by critical inflammatory cytokines TNF-α and IL-1β in human monocytic THP-1 cells. We optimized methods to combine two different approaches, bio-orthogonal non-canonical amino acid tagging (BONCAT) along with proteomics using isobaric tags (iTRAQ). BONCAT employed the incorporation of l-azidohomoalanine (AHA), an analog of methionine, into TNF-α or IL-1β induced nascent proteins. The AHA-containing nascent proteins were tagged with alkyne-biotin to allow enrichment using avidin affinity purification. The differential expressions of the enriched proteins were further determined using iTRAQ reagents and mass spectrometry (MS). The combination of BONCAT and proteomics represents a unique approach that has uncovered the nascent proteome induced by inflammatory cytokines TNF-α and IL-1β.
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| http://dx.doi.org/10.1016/j.jim.2012.06.003 | DOI Listing |
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