Abstract
Absolute quantification in proteomics usually involves simultaneous determination of representative proteolytic peptides and stable isotope–labeled analogs. The principal limitation to widespread implementation of this approach is the availability of standard signature peptides in accurately known amounts. We report the successful design and construction of an artificial gene encoding a concatenation of tryptic peptides (QCAT protein) from several chick (Gallus gallus) skeletal muscle proteins and features for quantification and purification.
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Acknowledgements
This work was supported by Biotechnology and Biological Sciences Research Council grants to R.J.B. and S.J.G. We are grateful to M. Fischer, Entelechon for his advice. We are grateful to B. Callen for assistance with expression of the QCAT.
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The QCAT concept has been submitted for patent protection by Entelechon and three of the authors (RJB, SJG and JMP) are cited as inventors.
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Supplementary Figure 1
The DNA sequence, translated protein sequence and features of the QCAT. (PDF 639 kb)
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Beynon, R., Doherty, M., Pratt, J. et al. Multiplexed absolute quantification in proteomics using artificial QCAT proteins of concatenated signature peptides. Nat Methods 2, 587–589 (2005). https://doi.org/10.1038/nmeth774
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DOI: https://doi.org/10.1038/nmeth774
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