Table 2 Overview of Nt-acetylation experimental techniques
COFRADIC | SILProNAQ | In vitro peptide library | In vitro NAT assay | NBD-Cl assay | |
|---|---|---|---|---|---|
Detects native acetylations? | Yes | Yes | No | No | Yes (in aggregate) |
Substrate discovery? | Yes | Yes | Yes | No | No |
Proteome scale or single substrate? | Proteome scale | Proteome scale | Many substrates, but it uses artificial N-termini | Single substrate | Proteome scale, but it does not distinguish individual substrates |
Cost | $$$$ | $$$ | $$ | $ | $ |
Can identify partially Nt-acetylated proteins? | Yes | Yes | No | No | No |
Applicable to patient samples / model organism tissues? | Yes | Yes | No | No | Yes |
Main advantages | Powerful, proteome-scale quantitation and substrate discovery, detects native acetylations, superior coverage | Powerful, proteome-scale quantitation and substrate discovery, detects native acetylations, less extensive fractionation compared to COFRADIC | Quantitative and unbiased substrate specificity discovery | Determines kinetic parameters, can be used for inhibitor studies, several readout options available (DTNB, HPLC, and 14C-Ac-CoA), suitable for high-throughput screening | Quick and simple, useful for a wide range of samples, detects native acetylations (in aggregate) |
Main drawbacks | Expensive and time-consuming, extensive fractionation compared to SILProNAQ | Expensive and time-consuming, low coverage compared to COFRADIC | Kinetics not possible | Single substrate only, relatively insensitive (depending on readout method) | No substrate discrimination |
Key references |
