Fig. 2: Schematic overview of QconCAT strategy. | Nature Communications

Fig. 2: Schematic overview of QconCAT strategy.

From: Decoding the stoichiometric composition and organisation of bacterial metabolosomes

Fig. 2: Schematic overview of QconCAT strategy.The alternative text for this image may have been generated using AI.

The QconCAT gene was subcloned into a plasmid vector (pEU01) optimised for cell-free system (CFS). The CFS was employed for transcription and translation to express the QconCAT proteins labelled with stable isotopes ([13C6,15N4]arginine and [13C6,15N2]lysine). The labelled QconCAT proteins (heavy, purple) were purified, quantified, and added to isolated Pdu MCPs from WT or ΔpduA cells with four replicated (unlabelled, light, green) in known quantity. After trypsin digestion, analyte mix released each of the QconCAT peptides in a strict stoichiometry of 1:1. LC-MS analysis allowed the quantification of each represented peptide of the isolated Pdu MCPs. One signature peptide for PduL, SVSQVEIS, was shown as an example.

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