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A cautionary note on the use of cap analogue affinity resins
- Cencic, Regina, Pelletier, Jerry
- Analytical biochemistry 2018 v.560 pp. 24-29
- affinity chromatography, messenger RNA, proteins, resins, ribosomes, translation (genetics)
- All cellular cytoplasmic mRNAs are capped at their 5’ ends with an m⁷GpppN group. Several proteins that mediate cap function have been identified by cap affinity purification, enabling their characterization in a number of biological processes. Among these, eukaryotic initiation factor (eIF) 4E is the best characterized and plays a critical role in regulating ribosome recruitment to mRNAs during translation initiation. Cap affinity chromatography is often used to identify eIF4E-interacting proteins, which could play critical roles in molding the eIF4E-interactome and impacting on eIF4E-directed translation. Here we address how improper implementation of this technology can lead to false conclusions and provide recommendations to ensure correct interpretation of data obtained by this approach.